NF-kappa B ( P65 or P65/P50 heterodimer ) bound to the P sequence in electrophoretic mobility shift assays ( EMSA ) and activated transcription through the P sequence when expression plasmids were cotransfected with P sequence-driven reporter plasmids in Jurkat T cells .

<protein>NF-kappa B</protein> ( <protein>P65</protein> or <protein>P65/P50 heterodimer</protein> ) bound to the <dna>P sequence</dna> in electrophoretic mobility shift assays ( EMSA ) and activated transcription through the <dna>P sequence</dna> when <dna>expression plasmids</dna> were cotransfected with <dna>P sequence-driven reporter plasmids</dna> in <cell_line>Jurkat T cells</cell_line> .


In EMSAs , NF ( P ) binding was inhibited by the unlabeled NF-AT binding site but not by the unlabeled AP1 binding site and purified NF-AT contained an activity that bound to the P sequence .

In EMSAs , <protein>NF ( P )</protein> binding was inhibited by the <dna>unlabeled NF-AT binding site</dna> but not by the <dna>unlabeled AP1 binding site</dna> and <protein>purified NF-AT</protein> contained an activity that bound to the <dna>P sequence</dna> .


Both mobility shift and sequence specificity of NF-AT were similar to those of NF ( P ) and only a small amount of P65 was detected in NF ( P ) in crude nuclear extracts .

Both mobility shift and sequence specificity of <protein>NF-AT</protein> were similar to those of <protein>NF ( P )</protein> and only a small amount of <protein>P65</protein> was detected in <protein>NF ( P )</protein> in crude nuclear extracts .


These results indicate that the component ( s ) of NF-AT has the potential to reconstitute NF ( P ) whereas NF-kappa B alone can not account for NF ( P ) in crude extracts .

These results indicate that the component ( s ) of <protein>NF-AT</protein> has the potential to reconstitute <protein>NF ( P )</protein> whereas <protein>NF-kappa B</protein> alone can not account for <protein>NF ( P )</protein> in crude extracts .


Unlike NF-AT , NF ( P ) does not contain AP1 as its DNA binding component .

Unlike <protein>NF-AT</protein> , <protein>NF ( P )</protein> does not contain <protein>AP1</protein> as its <protein>DNA binding component</protein> .


Activation of early growth response 1 gene transcription and pp90rsk during induction of monocytic differentiation .

Activation of <dna>early growth response 1 gene</dna> transcription and <protein>pp90rsk</protein> during induction of monocytic differentiation .


The present work has studied mechanisms responsible for induction of early growth response 1 ( EGR-1 ) gene expression during monocytic differentiation of U-937 myeloid leukemia cells .

The present work has studied mechanisms responsible for induction of <dna>early growth response 1 ( EGR-1 ) gene</dna> expression during monocytic differentiation of <cell_line>U-937 myeloid leukemia cells</cell_line> .


Differentiation of U-937 cells with 12-O-tetradecanoylphorbol-13-acetate ( TPA ) , an activator of the serine/threonine protein kinase C , was associated with transcriptional activation of EGR-1 promoter-reporter constructs .

Differentiation of <cell_line>U-937 cells</cell_line> with 12-O-tetradecanoylphorbol-13-acetate ( TPA ) , an activator of the <protein>serine/threonine protein kinase C</protein> , was associated with transcriptional activation of <dna>EGR-1 promoter-reporter constructs</dna> .


The EGR-1 promoter contains six CC ( A/T ) 6GG ( CArG ) motifs .

The <dna>EGR-1 promoter</dna> contains six CC ( A/T ) 6GG ( CArG ) motifs .


The two 5'-most distal CArG sequences conferred TPA inducibility .

The two <dna>5'-most distal CArG sequences</dna> conferred TPA inducibility .


In contrast , there was little effect of TPA on EGR-1 transcription in a TPA-resistant U-937 cell variant , designated TUR .

In contrast , there was little effect of TPA on <dna>EGR-1</dna> transcription in a <cell_line>TPA-resistant U-937 cell variant</cell_line> , designated <cell_line>TUR</cell_line> .


Treatment of both U-937 and TUR cells with okadaic acid , an inhibitor of serine/threonine protein phosphatases 1 and 2A , was associated with induction of monocytic differentiation and EGR-1 transcription through the 5'-most CArG element .

Treatment of both <cell_line>U-937</cell_line> and <cell_line>TUR cells</cell_line> with okadaic acid , an inhibitor of <protein>serine/threonine protein phosphatases 1 and 2A</protein> , was associated with induction of monocytic differentiation and <dna>EGR-1</dna> transcription through the <dna>5'-most CArG element</dna> .


Since these findings supported the involvement of serine/threonine protein phosphorylation in the regulation of EGR-1 expression , we studied activation of the 40S ribosomal protein S6 serine/threonine kinases , pp70S6K and pp90rsk .

Since these findings supported the involvement of serine/threonine protein phosphorylation in the regulation of <dna>EGR-1</dna> expression , we studied activation of the <protein>40S ribosomal protein S6 serine/threonine kinases</protein> , <protein>pp70S6K</protein> and <protein>pp90rsk</protein> .


Although both kinases participate in regulating cell growth , there was no detectable activation of pp70S6K during TPA- or okadaic acid-induced monocytic differentiation .

Although both kinases participate in regulating cell growth , there was no detectable activation of <protein>pp70S6K</protein> during TPA- or okadaic acid-induced monocytic differentiation .


Moreover , rapamycin , an inhibitor of pp70S6K activation , had no effect on induction of EGR-1 expression .

Moreover , rapamycin , an inhibitor of <protein>pp70S6K</protein> activation , had no effect on induction of <dna>EGR-1</dna> expression .


In contrast , analysis of pp90rsk activity by phosphorylation of a peptide derived from S6 protein demonstrated stimulation of this kinase in TPA-treated U-937 , and not TUR , cells .

In contrast , analysis of <protein>pp90rsk</protein> activity by phosphorylation of a peptide derived from <protein>S6 protein</protein> demonstrated stimulation of this <protein>kinase</protein> in <cell_line>TPA-treated U-937 , and not TUR , cells</cell_line> .


Okadaic acid treatment of both cell types was associated with activation of pp90rsk .

Okadaic acid treatment of both cell types was associated with activation of <protein>pp90rsk</protein> .


Antioxidants inhibit monocyte adhesion by suppressing nuclear factor-kappa B mobilization and induction of vascular cell adhesion molecule-1 in endothelial cells stimulated to generate radicals .

Antioxidants inhibit <cell_type>monocyte</cell_type> adhesion by suppressing <protein>nuclear factor-kappa B</protein> mobilization and induction of <protein>vascular cell adhesion molecule-1</protein> in <cell_type>endothelial cells</cell_type> stimulated to generate radicals .


Cell adhesion to endothelial cells stimulated by tumor necrosis factor-alpha ( TNF ) is due to induction of surface receptors , such as vascular cell adhesion molecule-1 ( VCAM-1 ) .

Cell adhesion to <cell_type>endothelial cells</cell_type> <protein>stimulated by tumor necrosis factor-alpha</protein> ( <protein>TNF</protein> ) is due to induction of <protein>surface receptors</protein> , such as <protein>vascular cell adhesion molecule-1</protein> ( <protein>VCAM-1</protein> ) .


The antioxidant pyrrolidine dithiocarbamate ( PDTC ) specifically inhibits activation of nuclear factor-kappa B ( NF-kappa B ) .

The antioxidant pyrrolidine dithiocarbamate ( PDTC ) specifically inhibits activation of <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappa B</protein> ) .


Since kappa B motifs are present in VCAM-1 and intercellular adhesion molecule-1 ( ICAM-1 ) promoters , we used PDTC to study the regulatory mechanisms of VCAM-1 and ICAM-1 induction and subsequent monocyte adhesion in TNF-treated human umbilical vein endothelial cells ( HUVECs ) .

Since <dna>kappa B motifs</dna> are present in <dna>VCAM-1 and intercellular adhesion molecule-1 ( ICAM-1 ) promoters</dna> , we used PDTC to study the regulatory mechanisms of <protein>VCAM-1</protein> and <protein>ICAM-1</protein> induction and subsequent <cell_type>monocyte</cell_type> adhesion in <cell_line>TNF-treated human umbilical vein endothelial cells</cell_line> ( <cell_line>HUVECs</cell_line> ) .


PDTC or N-acetylcysteine dose dependently reduced TNF-induced VCAM-1 but not ICAM-1 surface protein ( also in human umbilical arterial endothelial cells ) and mRNA expression ( by 70 % at 100 mumol/L PDTC ) in HUVECs as assessed by flow cytometry and polymerase chain reaction .

PDTC or N-acetylcysteine dose dependently reduced <protein>TNF-induced VCAM-1</protein> but not <protein>ICAM-1 surface protein</protein> ( also in <cell_type>human umbilical arterial endothelial cells</cell_type> ) and mRNA expression ( by 70 % at 100 mumol/L PDTC ) in <cell_line>HUVECs</cell_line> as assessed by flow cytometry and polymerase chain reaction .


Gel-shift analysis in HUVECs demonstrated that PDTC prevented NF-kappa B mobilization by TNF , suggesting that only VCAM-1 induction was controlled by NF-kappa B .

Gel-shift analysis in <cell_line>HUVECs</cell_line> demonstrated that PDTC prevented <protein>NF-kappa B</protein> mobilization by <protein>TNF</protein> , suggesting that only <protein>VCAM-1</protein> induction was controlled by <protein>NF-kappa B</protein> .