Figure 8

HDACs are not a major target for SENP1 action on PR transcriptional activity. A) TSA enhances PR-B protein stability. HeLa cells were transiently transfected with expression vectors encoding wild type PR-B. Cells were treated 24 hrs without (-) or with (+) 10 nM R5020 in the presence of increasing concentration of TSA. Western blot analysis was performed on cell extracts probed with the anti-PR1294 monoclonal and anti β-actin antibodies. B) HeLa cells were transfected with 2 μg of PRE2-luciferase reporters together with 50 ng of a PR-B (left), or the PR-B K388R mutant (right) expression vectors and Renilla-Luc as an internal control in the presence or absence of 100 ng SENP1 expression vectors. The cells were treated for 24 hrs with the agonist R5020 (10 nM), without (-) or with (+) 100 nM TSA then harvested and lysed. The extracts were assayed for luciferase activities as in Figure 1. Statistical significance was computed by unpaired student's t test. *p < 0.05.