Fig. 3.

Endogenous HEXIM1 Negatively Modulates Glucocorticoid-Mediated Transcriptional Activation

A, Cell lysates from various tissues from adult mice were subjected to Western blot analysis using indicated antibodies. B, HepG2 cells were infected with Adsictrl or AdsiHEXIM1 in phenol red-free Opti-MEM I at MOI of 100 for indicated time periods. Whole-cell extracts were prepared, and protein expression levels of endogenous HEXIM1, GR, and α-actinin were assessed in Western blotting. C, HepG2 cells were infected with the recombinant adenoviruses for 36 h as described in panel B and stimulated with 100 nm DEX for 6 h. Total RNA was prepared and mRNA for atp1a1, scnn1a, and gapdh was measured with qRT-PCR. Samples were normalized to gapdh, and relative expression levels to vehicle-treated samples are presented as relative mRNA expression. Error bars represent sd values of at least three independent experiments. D, HepG2 cells were infected with the recombinant adenoviruses for 36 h as described in panel B and stimulated with 1 μm DEX for indicated time periods. ChIP assays were performed with the indicated antibodies as described in Materials and Methods. IP, Immunoprecipitation.