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TPL induced phosphorylation of Rpb1 by activation of protein kinases.

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posted on 20.02.2013, 14:52 by Ying Wang, Jin-jian Lu, Li He, Qiang Yu

(A) Comparison of the effect of TPL with NaF on Rpb1. Hela cells were treated with TPL or NaF for 3 h in the presence/absence of MG132. Whole cell lysates were processed for SDS-PAGE and Western blot analysis using antibodies as indicated. (B) Hela cells were pretreated with MG132 for 1 h, and then treated with TPL in the presence/absence of 50 µM H7 for 3 h. Whole cell lysates were subjected to SDS-PAGE and Western blot analysis. (C) Hela cells were pretreated with MG132 for 1 h, and then treated with TPL in the presence/absence of 50 µM DRB for 3 h. Whole cell lysates were subjected to SDS-PAGE and Western blot analysis. (D) Analysis of effect of CDK9 and Cyclin T1 siRNA on TPL-induced Rpb1 phosphorylation. Hela cells were transfected with NC (negative control) or CDK9 and Cyclin T1 siRNA. 48 h after transfection, the Hela cells were pretreated with 20 µM MG132 for 1 h. Then the cells were treated with DMSO or 125 nM TPL for another 3 h before collected to SDS-PAGE and Western blot analysis. β-Actin served as a loading control.

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