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PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
Next, we applied two different analyses to measure cell death.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
We further analyzed the FACS data, particularly cell populations at sub-G1 phase, in which cells undergo cell death.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
This cell population contained both floating cells in culture media and cells attached to the culture plate.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
We found additive effects on cell death indices in combined treatment groups with KCC-07 and DNA damaging reagents in both cell lines (Fig. 3B).
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
Additionally, dead cells were measured in real-time by live cell imaging using IncuCyte.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
However, the dead cell index was not significantly different between cell lines with DNA damage induction and combined treatment (Fig. 3C), most likely due to measurement of only the attached cell population on the plate using IncuCyte.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
Of note, SH-SY5Y cell lines showed aggregation of cells upon KCC-07 treatment, which was also detected in combined treatment groups, indicating that cell response to KCC-07 might vary depending on cell lines.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
Moreover, we measured proliferation indices in continuous exposure to KCC-07 and/or DNA damaging reagents.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
There was further proliferation reduction in U-87MG and SH-SY5Y cell lines with combined treatment of KCC-07 and DNA damaging reagents compared to individual treatments (Fig. 4A), suggesting that additive effects on growth suppression of tumor cells could be achieved by combined treatment with KCC-07 and DNA damage induction.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
Collectively, these data demonstrate the expandable application of KCC-07 to other neural tumors and its potential to enhance the efficacy of existing treatments.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
The combination approach with KCC-07 suggested here could enable dose reduction of DNA damaging agents, substantially mitigating their associated toxicity while preserving or even enhancing therapeutic efficacy for brain tumors.
PMC12426421
KCC-07, MBD2 Inhibitor, Expands the Therapeutic Window of DNA Damage Inducing Reagents in Neural Tumor Cells
In addition, further studies are warranted to identify the specific mediators that bridge MBD2 inhibition and p53 pathway activation in U-87MG and SH-SY5Y cell lines, which could provide valuable insights for optimizing KCC-07 based therapeutic strategies in neural tumors.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Background and Objectives: Neuroblastoma is the most common extracranial solid tumor in children, often presenting challenges in treatment due to its clinical and genetic heterogeneity.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
This study investigated the anticancer potential of Pelargonium sidoides root extract on the human neuroblastoma cell line (SH-SY5Y).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Using XTT assays, ELISA-based oxidative stress markers, and RT-PCR analysis of apoptotic genes, the study explored the extract’s effects on cell proliferation, oxidative stress, and apoptosis.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Materials and Methods: For the cell culture, SH-SY5Y human neuroblastoma cells were thawed, cultured, and maintained under appropriate conditions for experiments.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The dose- and time-dependent activity of Pelorgonium sidoides extract on SH-SY5Y neuroblastoma cells was investigated by XTT assay.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The change in the oxidative stress marker 8-Hydroxy-2′-deoxyguanosine (8-OhDG) level was determined by ELISA for the doses applied to the control group root extract at a concentration of 25 μg/mL. Total antioxidant status (TAS) and total oxidant status (TOS) were measured from the cells in the study group with the help of a commercial kit.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The oxidative stress index (OSI) was calculated by dividing the TAS by the TOS and multiplying by 100.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
In order to evaluate the expression levels of apoptosis-related Bax, Bcl-2, Caspase-3, Caspase-8, and Caspase-9 genes at the mRNA level in control and dose group cells, RNA isolation was performed from the SH-SY5Y control and dose group cells (IC50 value).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Results: It is observed that the P. sidoides substance inhibits proliferation in cells at 24 h (p < 0.05).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
As the dose increases, cell proliferation decreases (p < 0.05).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The IC50 value was calculated to be 113.83 μg/mL at 24 h. The concentration of 8-OhDG increased in neuroblastoma cells as a result of P. sidoides extract treatment (p < 0.05).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
TOS levels increased in neuroblastoma cells treated with P. sidoides extract (p < 0.01).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
OSI levels increased in cells treated with P. sidoides extract (p < 0.001).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
BAX and Caspase-8 expression increased are statistically significant in the P. sidoides dose group (p < 0.05).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Conclusions: P. sidoides extract induces apoptosis in neuroblastoma cells through oxidative stress and mitochondrial- and death receptor-mediated pathways.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
This study highlights the potential of P. sidoides as a complementary therapeutic agent for neuroblastoma, warranting further in vivo and clinical investigations to assess its safety and efficacy.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Childhood cancers stand out as a serious public health problem in the medical field.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Every year, thousands of children are diagnosed with cancer worldwide, and these diseases are responsible for a significant portion of childhood deaths .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Early diagnosis and treatment of these cancers are of great importance for both the course of the disease and the quality of life of patients.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Cancers seen in childhood generally originate from cells with high proliferative potential, and in addition to genetic factors, environmental effects also play a role in the emergence of these diseases .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Among childhood cancers, neuroblastoma holds a particularly critical position.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
It is the most common extracranial solid tumor diagnosed in children and accounts for approximately 15% of all pediatric cancer-related deaths .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Originating from primitive neural crest cells in the embryonic period, this tumor is the third most common type of cancer in childhood, with a rate of 7% .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Neuroblastoma, which shows great clinical and genetic heterogeneity, presents challenges that limit the effectiveness of current treatment methods .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The pathogenesis of neuroblastoma involves a complex interplay of genetic, epigenetic, and environmental factors.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Approximately 90% of neuroblastoma patients exhibit chromosomal changes, including MYCN amplification, ALK mutations, and 1p or 11q deletions, which are linked to disease progression and poor outcomes .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
These alterations underscore the need for targeted therapies that can address these genetic aberrations.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Current treatment strategies for neuroblastoma typically include surgery, chemotherapy, radiotherapy, and immunotherapy.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
However, these approaches often come with significant toxicity, particularly in young patients, and are limited in their efficacy for high-risk cases.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
As such, there is an urgent need for novel therapeutic strategies that can improve survival while minimizing side effects.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
In recent years, intensive studies have been conducted on the role of plant-based bioactive molecules in cancer treatment .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
It has been shown that these molecules have the potential to suppress tumor proliferation, activate apoptosis mechanisms, and regulate oxidative stress levels .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Pelargonium sidoides (P. sidoides) root extract has been used in traditional treatment methods for a long time and has been found to be effective in modern medicine, especially in the treatment of upper respiratory tract infections .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
However, recent studies also focus on the potential anticancer effects of P. sidoides and show that this plant may affect various biological pathways associated with cancer .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The phenolic compounds of P. sidoides attract attention with their potential to regulate PI3K-Akt and Ras/MAPK pathways, which are cancer-related signaling pathways .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The aim of this study was to investigate the effects of P. sidoides root extract on the SH-SY5Y human neuroblastoma cell line.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The effects of the extract on cell proliferation, oxidative stress levels and apoptosis mechanisms were examined with various experimental methods, and the findings are presented in a way that will contribute to new treatment approaches for childhood cancers.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
This study was supported by the Ordu University scientific research project (BAP project number: A-2308).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The study was conducted between 6 June 2023, and 29 May 2024.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
For the cell culture, SH-SY5Y human neuroblastoma cells were thawed, cultured, and maintained under appropriate conditions for experiments.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Cell proliferation was achieved using DMEM-F12 culture medium containing 10% fetal bovine serum, 2 mM L-glutamine, and 1% penicillin–streptomycin in a 95% humidity and 5% CO2 oven at 37 °C.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Cells were multiplied under appropriate conditions and made ready for experiments.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
In order to renew our stocks, they were frozen in new cryos with DMSO and stored at minus 80 °C.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The dose- and time-dependent activity of Pelorgonium sidoides extract on SH-SY5Y neuroblastoma cells was investigated by XTT assay.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Briefly, cells plated in 96-well plates were treated with the synthesized substance at appropriate dose ranges specified in the literature for 24, 48, and 72 h, following an initial incubation period of 24 h. After incubation, XTT solution was added to the 96-well plates, and after 4 h, absorbance values (OD) were measured using an ELISA reader at a wavelength of 450 nm with a reference range of 630 nm.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The percentage of cell viability was calculated by dividing the optical density value measured in each well by the control optical density value and multiplying by one hundred.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, non-toxic doses were determined, and dose selection was made for further experiments.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The inhibitory concentration (IC50) value was calculated via the (https://www.aatbio.com/tools/ic50-calculator, accessed on 20 December 2023). %
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Cell viability = [(measured optical density value)/(control optical density value)] × 100.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The change in the oxidative stress marker 8-Hydroxy-2′-deoxyguanosine level was determined by ELISA for the doses applied to the control group root extract at a concentration of 25 μg/mL. A human 8-Hydroxy-deoxyguanosine ELISA kit (Cat No: EA0048Hu; Bioassay Technology Laboratory, Shanghai, China) was used.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The application procedure was as follows: First, the standard concentration provided with the kit was prepared by serial dilution.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Standard concentrations were adjusted to be 800, 400, 200, 100, and 50 ng/mL from the main stock of 1600 ng/mL. Then, 20 mL of wash solution provided as 30X was diluted to 1X with distilled water.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Kit components were brought to room temperature, and the study began.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Standards were loaded into 96-well ELISA plates as 50 μL. No antibody was added to the standards.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Samples were loaded as 40 μL and then 10 μL of anti-8-OHDG antibody was added.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, 50 μL of streptavidin–HRP was added to both standards and samples.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
This addition was not made for the blank well.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
After all these added components, incubation was applied for 60 min at 37 °C in the dark.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
At the end of the incubation, washing was performed 5 times for all wells with wash solution.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, substrate solution A and 50 μL of substrate solution B were pipetted.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, incubation was performed for 10 min at 37 °C in the dark.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
After the incubation was completed, 50 μL of stop solution was added and readings were performed using a microplate reader (Epoch 2) at 450 nm within 10 min.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
A similar protocol was applied for all other ELISA tests.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The principle of total oxidant level measurement is based on the conversion of the ferrous ion chelator complex of the oxidants in the sample to ferric ion, which reacts with the chromogen in an acidic environment and causes an increase in absorbance.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The increase in absorbance observed spectrophotometrically is directly proportional to the oxidant molecules in the sample .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The principle of total antioxidant level measurement is based on the principle that all antioxidants in the sample convert the blue–green ABTS radical into colorless reduced ABTS .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
In this study, a control and dose group were created for SH-SY5Y neuroblastoma cells in order to investigate the effect of P. sidoides root extract.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Total oxidant and total antioxidant levels were measured from the cells in the study group with the help of the commercial kit provided (TAS and TOS, Rel Assay, Gaziantep, Turkey.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
OSI was calculated by dividing the TAS by the TOS and multiplying by 100.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The experiments were repeated three times to ensure the reliability of the experiments .
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
In order to evaluate the expression levels of apoptosis-related Bax, Bcl-2, Caspase-3, Caspase-8, and Caspase-9 genes at the mRNA level in control and dose group cells, RNA isolation was performed from the SH-SY5Y control and dose group cells (IC50 value).
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Neuroblastoma cells were plated in 6-well cell culture plates at a density of 1 × 10 to 2 × 10 cells per well and incubated for 24 h to achieve appropriate confluency for further experiments.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, control and dose groups were adjusted.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
At the end of incubation, the medium in the wells was removed and the cells were washed with 3 mL cold PBS.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
After removing the PBS, 500 μL Trizol was added to the wells to lift the cells.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The homogenate was transferred to 1.7 mL Eppendorf tubes and incubated for 10 min at room temperature.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, 100 μL of chloroform was added to each Eppendorf tube and vortexed, and incubated again at room temperature for 15 min.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Then, the tubes were centrifuged at 15,000× g for 15 min at 4 °C, and the colorless upper phase containing RNA was collected and transferred into separate Eppendorf tubes.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
A total of 250 μL of isopropanol was added to the collected upper phase, pipetted, and left at room temperature for 10 min.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
After the tubes were centrifuged at 15,000× g for 10 min at +40 °C, the supernatant was carefully discarded, and 70% ethanol was added to the pellet and centrifuged at 12,000× g for 10 min at +40 °C.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The supernatant then was discarded again and the pellet was air dried for a short time.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
Finally, the pellet was dissolved with 40 μL of RNase–DNase-free water.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
The quantity and quality of the obtained RNAs were determined using Nanodrop.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
A260/A280 and A260/A230 ratios were evaluated to determine the phenol, protein, and genomic DNA contamination that may occur during isolation.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
After UV measurements, RNA samples with 2 ± 0.1 for A260/A280 and 2.0–2.4 for A260/A230 were used in the analyses.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
For mRNA expression analysis, cDNA synthesis was performed from isolated RNAs using the A.B.T.™ cDNA Synthesis Kit with RNase Inhibitor (High-Capacity) synthesis kit (CatNo: C03-01-20, ABT, Ankara, Turkey) along with a random hexamer, dNTP, and reverse transcriptase enzyme (RT) followingg the manufacturer’s protocol.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
After the mixture was prepared, it was incubated at 25 °C for 10 minutes and then at 37 °C for 120 minutes for cDNA synthesis.
PMC11679892
Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells
At the end of this period, the mixturewas kept at 85 °C for 5 min to inhibit the enzyme.