PMCID string | Title string | Sentences string |
|---|---|---|
PMC10728200 | Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination | STUB1 overexpression promotes the ubiquitination of GPX4, thereby inducing ferroptosis in gastrointestinal stromal tumors (GIST). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Despite targeted therapy with tyrosine kinase inhibitors (TKIs), unresectable or metastatic human GISTs generally relapse under treatment. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Thus, alternative therapeutic approaches are needed to overcome GIST resistance. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Ferroptosis, a cell death driven by iron-dependent phospholipid peroxidation, is an emerging therapeutic approach in cancers. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | However, the potential of ferroptosis induction in GISTs remains largely unknown. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | This study shows that GIST cell lines are highly sensitive to the type II ferroptosis inducer, RSL3. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We also provide evidence that inhibition of YAP by verteporfin (VP) promotes ferroptosis in GIST cells while, surprisingly, CA3 mediates ferroptosis independently of YAP inhibition. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Finally, we highlight a positive correlation between the highly expressed transferrin receptor protein 1 (TFRC), GISTs with elevated mitotic counts or higher risk, and with YAP expression/activation in human GIST tissue microarrays (TMA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Taken together, our results suggest that induction of ferroptosis and/or modulation of YAP activity offer promising perspectives for GIST treatment. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | GISTs are sarcomas of the gastrointestinal tract often associated with gain-of-function mutations in KIT or PDGFRA receptor genes. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | While most GISTs initially respond to tyrosine kinase inhibitors, relapses due to acquired resistance frequently occur. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The induction of ferroptosis, an iron-dependent form of non-apoptotic cell death, emerged as a novel therapeutic approach in cancers and remains poorly characterized in GISTs. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We studied hallmarks of ferroptosis, i.e., lipid peroxidation, iron and glutathione content, and GPX4 protein expression in imatinib-sensitive (GIST882) and -resistant (GIST48) GIST cell lines. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | GIST cells were highly sensitive to the induction of ferroptosis by RSL3, which was reversed by liproxstatin and deferoxamine. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Lipid peroxidation and ferroptosis were mediated by VP and CA3 in GIST cells through a significant decrease in antioxidant defenses. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Moreover, VP, but surprisingly not CA3, inhibited a series of target genes downstream of YAP in GIST cells. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The ferroptosis marker TFRC was also investigated by immunohistochemistry in GIST tissue arrays. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | TFRC expression was observed in all samples. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | High TFRC expression was positively correlated with high-risk GISTs, elevated mitotic count, and YAP nuclear localization, reflecting YAP activation. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | This study highlights ferroptosis as a novel cell death mechanism in GISTs, and a potential therapeutic target to overcome resistance to tyrosine kinase inhibitors. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Gastrointestinal stromal tumors (GISTs) are the most frequent sarcomas of the gastrointestinal tract and derive from the interstitial cells of Cajal (ICCs) or their precursors . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | ICCs are key players in the motility of the gut as they act as a pacemaker of the muscularis propria and mediate the transduction of the enteric nervous system inputs . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Approximately 85% of GISTs are driven by somatic gain-of-function mutations of the gene encoding the tyrosine kinase receptor (RTK) KIT (CD117) . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Surgery is the only curative treatment for GISTs; however, for unresectable or metastatic disease, the tyrosine kinase inhibitor (TKI) imatinib (Gleevec, Glivec) is the first line of therapy . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Unfortunately, most patients ultimately progress due to primary or secondary mutations of KIT or PDGFRA. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Second- and third-line TKIs (regorafenib, sunitinib, etc.) |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | provide only limited benefits . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Therefore, the development of novel therapeutic strategies remains essential to improve patient outcome. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We previously reported the involvement of the transcriptional co-activator YAP in the survival of imatinib-sensitive (GIST882) and -resistant (GIST48) human GIST cell lines . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | YAP is located downstream of the Hippo pathway, which responds to diverse stimuli including cell–cell interaction, cellular stress, extracellular signals, cell polarity, and mechanical signals . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | In its active unphosphorylated state, YAP binds transcriptional cofactors such as TEA domain family members (TEADs), and activates a transcriptional program of genes involved in cell proliferation, survival, differentiation, and tumorigenesis . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The YAP inhibitor verteporfin (VP) elicited massive cytotoxicity in GIST882 and GIST48 cell lines . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Moreover, nuclear YAP immunoreactivity, indicative of YAP transcriptional activity, was observed in 71% of primary human GIST tissues, providing evidence of YAP involvement in GIST biology . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Recently, a series of links between the Hippo–YAP pathway and ferroptosis have been identified . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Ferroptosis is a non-apoptotic iron-dependent form of cell death characterized by an accumulation of lipid peroxidation that leads to membrane damage . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Ferroptosis is the result of an imbalance between oxidative damage and antioxidant defense . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Oxidative damage is either caused by an accumulation of radical oxygen species (ROS), mainly produced by the Fenton reaction which involves free ferrous iron and H2O2, or lipid peroxidation mediated by iron-containing lipoxygenases (LOX) . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Free ferrous iron is mainly represented by the labile iron pool (LIP), which is regulated by different proteins involved in iron metabolism, such as transferrin receptor 1 (TFRC), ferritin (FTH, FTL), and ferroportin (FPN1). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The antioxidant defenses involve the axis between SLC7A11, glutathione (GSH), glutathione peroxidase 4 (GPX4), and the nuclear factor erythroid 2-related factor 2 (NRF2). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | SLC7A11 is one of the two subunits of the cystine–glutamate antiporter system Xc that imports extracellular cystine into the cytoplasm, where it is used in the biosynthesis of GSH, the cofactor of the antioxidant enzyme GPX4. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | This later reduces hydroperoxide lipids and thus prevents ferroptosis . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | As ferroptosis emerges as a promising approach for cancer therapy , it appeared as an attractive mechanism to explore in GISTs. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Ferroptosis sensitivity has recently been reported in imatinib-sensitive GIST-T1 cells (KIT exon 11 mutation ) and imatinib-resistant GIST R8 cells , which have been established from parental GIST-T1 cells treated with increasing doses of imatinib . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | However, so far, ferroptosis has not been investigated in imatinib-sensitive GIST882 (KIT exon 13 mutation) cells, or in imatinib-resistant GIST48 cells (KIT primary exon 11 and resistant exon 17 mutations), which originate from a GIST patient who had progressed under imatinib treatment. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Here, we studied the impact of the class II ferroptosis inducer (FIN), RSL3, a direct GPX4 inhibitor, on the viability and lipid peroxidation of GIST882 and GIST48 cells. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We next used two YAP inhibitors and tested different hallmarks of ferroptosis in GIST882 and GIST48 cells. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Finally, as cancer cells appear to be “addicted to iron” and as the TFRC emerged as a new cancer marker , we tested the expression of the TFRC in human GIST tissues and explored possible correlations with clinicopathological criteria and YAP expression. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The collection and analysis of human tissue samples were conducted in line with ethical standards according to the Declaration of Helsinki, as well as national and international guidelines. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The study was approved by the Institutional Medical Ethics Committees of Erasmus Hospital and Faculty of Medicine, Université Libre de Bruxelles (reference number P2016/316); the Ethical Committee of Institut Jules Bordet (reference number CE 2964), Brussels, Belgium; and the Medical Ethics Committee, UZ Leuven, Leuven, Belgium (reference number S66100) for their respective human materials. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The human GIST882 cell line was kindly provided by Dr. Jonathan A. Fletcher, Harvard Medical School, Boston, MA, USA. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were cultured at 37 °C in DMEM (Gibco, CA, USA, Cat# 41965-062), supplemented with 10% (v/v) FBS, 2% (v/v) penicillin–streptomycin (Gibco, CA, USA, Cat# 15140-122). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The human GIST48 cell line was kindly provided by Dr. Ronald DeMatteo, Perelman School of Medicine, Philadelphia, PA, USA. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were cultured at 37 °C in RPMI-1640 (Gibco, CA, USA, Cat# 31870-074), supplemented with 10% (v/v) FBS, 1% (v/v) L-glutamine (Gibco, CA, USA, Cat# 35050-038), 1% (v/v) HEPES (Gibco, CA, USA, Cat# 15630-056), 0.1% (v/v) 2-mercaptoethanol (Gibco, CA, USA, Cat# 31350-010), and 0.5% (v/v) penicillin–streptomycin. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Verteporfin and cumene hydroperoxide were purchased from Sigma, St. Louis, MO, USA (Cat# C0737, SML0534, and 247502, respectively). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Erastin, liproxstatin-1, deferoxamine mesylate, RSL3, CA3, and staurosporine were purchased from Selleckchem, Houston, TX, USA (Cat# S7242, S7699, S5742, S8155, S8661, and S1421, respectively). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cell viability was estimated using a WST-1 assay (Roche, Indianapolis, IN, USA, Cat# 5015944001). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | GIST882 and GIST48 cells were seeded in 96-well plates (TPP Techno Plastic Products AG, Trasadingen, Switzerland) at a concentration of 10,000 cells/well supplemented with 100 μL of medium 72 h before drug treatments. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Drug concentrations were chosen based on previous studies . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | After drug treatment, 10 μL of WST-1 reagent was added and plates were incubated at 37 °C for either 2 h or 3 h for GIST882 and GIST48 cells, respectively. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Absorbance was measured at 450 nm on a plate reader (iMark Microplate Absorbance Reader, BioRad, Hercules, CA, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | IC50s were calculated using Prism 7 software (GraphPad Software, Inc., La Jolla, CA, USA) and CompuSyn software (https://www.combosyn.com/, accessed on 18 August 2022) Cohorts of archival GIST primary tumors and metastatic GISTs from formalin-fixed slides, paraffin-embedded (FFPE) material, and tissue microarrays (TMA) were obtained from the Laboratory of Pathological Anatomy, Jules Bordet Institute, Brussels, Belgium and from the Laboratory of Experimental Oncology, Department of Oncology, KU Leuven, Leuven, Belgium. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Clinicopathological features are given in Tables S1–S3. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The risk of relapse after the curative resection of primary GISTs was determined according to Miettinen’s classification . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | FFPE slides and TMA were rehydrated through toluol and graded alcohol solutions, before then being heated at 90 °C in citrate buffer 0.01M (pH 6.0) antigen retrieval solution for 20 min to unmask the antigen. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Slides were then cooled for 15 min before being put in a 0.1% H2O2–methanol solution for 30 min to block endogenous peroxidase. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | After washing, slides were incubated with a TBS-Triton X-100, 10% NHS blocking solution for 1 h, followed by the primary antibodies diluted in a TBS-Triton X-100 0.1% and 1% NHS solution overnight at RT in a humid chamber. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Sections were rinsed and incubated with a secondary biotinylated antibody for 1 h followed by an ABC solution (ABC Kit Standard; Vector Laboratories, Burlingame, CA, USA, Cat# PK-6100) for 1 h. Revelations with nickel-enhanced DAB (DAB-Ni) were performed at RT for 5–10 min, resulting in a black precipitate. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The DAB-Ni solution was prepared by dissolving 0.06 g of nickel ammonium sulfate (Fluka, Buchs, Switzerland, Cat# 09885) and 2 mg of DAB (Sigma-Aldrich, St. Louis, MO, USA, Cat# D5637) in 10 mL of 0.05 M Tris/HCl, pH 8. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Before use, 1 μL of 30% H2O2 (Thermo Fisher Scientific, MA, United States, Cat# 241022500) was added to the DAB-Ni solution. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Slides were dehydrated through graded alcohol and toluol solutions, mounted with DPX (Merck, NY, United States) and stored at room temperature. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | A list of antibodies is given in Table S4. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | For each TMA, KIT, YAP, and TFRC, staining intensities were scored individually by two researchers (Jean-Marie Vanderwinden and Marine Delvaux) blinded for clinicopathological information, and the results were assembled by consensus. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | For Bordet TMA, ten replicate samples were available, while for KU Leuven TMA, two replicate samples were available. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | For KIT and YAP staining, samples were scored as negative (−), positive (+), or strongly positive (++). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | None of the samples were graded as negative for KIT expression; thus, the expression scoring was +/++. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The YAP location was also evaluated and defined as nuclear (N) or diffuse (D) or mixed (M). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Nuclear and mixed YAP corresponded to active YAP, while diffuse YAP was considered to be an inactive YAP. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | TFRC staining was graded as negative (−), weakly positive (+), moderately positive (++), and strongly positive (+++). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | None of the samples were graded as negative for TFRC expression; thus, the TFRC expression grading was +/++/+++. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Image acquisition and analysis were performed at LiMiF https://limif.ulb.be/, the Light Microscopy Facility, Université Libre de Bruxelles, Faculty of Medicine, Campus Erasme, Brussels. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | IHC slides were observed on a AxioObserver Z1 inverted microscope (Zeiss, Jena, Germany), using a Plan Apochromat 20x/0.8 dry objective (Zeiss). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Transmitted light illumination was provided by a HAL100 halogen lamp and condenser in “bright-field position”. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Images (1920 by 1216 pixels, pixel size (x-y): 0.293 micron by 0.293 micron) were acquired with an Axiocam 702 monochrome camera (Zeiss) as proprietary *.czi files. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Files were processed with Zen 2.5 (Blue Edition) software (Zeiss). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Images were displayed in the linear mode with manual contrast adjustment and exported as 16 bits uncompressed TIF files. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The total RNA was extracted using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA, Cat# 74104), according to the manufacturer’s instructions. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Genomic DNA was removed using the RNase-Free DNase set (Qiagen, Valencia, CA, USA, Cat# 79274). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Then, 1 µg of RNA was retrotranscripted into cDNA using LunaScript RT SuperMix Kit (New England BioLab, Ipswich, MA, USA, Cat# E3010L), according to the manufacturer’s instructions. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The cDNA reverse transcription products were amplified with specific primers (Table S5) by qPCR using SYBR Green chemistry on a QuantStudio™ 3 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Identical thermal profile conditions, namely 95 °C for 10 min, followed by 40 cycles of 95 °C for 15 s and 60 °C for 1 min, were used for all primer sets. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Emitted fluorescence was measured during the annealing–extension phase and amplification plots were generated using the sequence detection system. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Transcriptional quantification, relative to GAPDH and HPRT1 reference genes, was performed using qBase+ software (Biogazelle, Zwijnaarde, Belgium). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Statistical analysis was performed with Prism 9 software (GraphPad Software, Inc., La Jolla, CA, USA), using the multiple-ratio paired t-test. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | RNA quality was checked with a fragment analyzer (Agilent Technologies). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Indexed cDNA libraries were obtained using the TruSeq Stranded mRNA Sample Prep Kit (Illumina), following the manufacturer’s recommendations. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The multiplexed libraries were loaded on a NovaSeq 6000 (Illumina) using a S2 flow cell and sequences were produced using a 200 Cycles Reagent Kit. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Paired-end reads were mapped against the human reference genome GRCh38 using STAR_2.5.3a software to generate read alignments for each sample. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Annotations of Homo_sapiens. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | GRCh38.90.gft were obtained from http://sftp.ensembl.org./ (accessed on 12 August 2022). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | After the transcripts were assembled, gene-level counts were obtained using HTSeq-0.9.1 and normalized to 20 million aligned reads. |
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