Fig. 4.

GR and HEXIM1 Stochastically Contribute to Hormone-Dependent Transcriptional Regulation of GR-Target Genes

A, HepG2 cells were infected with the recombinant adenoviruses expressing FLAG-GR and HEXIM1 in phenol red-free Opti-MEM I at MOI of 5 for 24 h as indicated. Cells were lysed and subjected to Western blot analysis using the indicated antibodies. B, HepG2 cells were infected with the recombinant adenoviruses as described in panel A and treated with or without 100 nm DEX for 6 h. Total RNA was prepared, and mRNA for atp1a1, sgk1, 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. C, HepG2 cells were infected with the recombinant adenoviruses in phenol red-free Opti-MEM I for 24 h as indicated and treated with or without 1 μm DEX for 20 min. ChIP assays were performed with anti-GR antibodies as described in Materials and Methods. IP, Immunoprecipitation.