FIG. 2.

Analysis of MMTV-array cell lines expressing GFP-GR truncation mutants. (A) Tetracycline-regulated expression of GFP-GR constructs. Total cell extracts from cells grown in the presence or absence of tetracycline were probed by Western blotting for the expression of the GFP-GR constructs with anti-GFP and anti-GR antibodies. Equal loading was verified by anti-actin antibody. (B) Transcriptional activity of GFP-GR-, GFP-407C-, or GFP-N525-expressing cells treated as indicated with either vehicle (−), 100 nM Dex, 1 μM Dex-Mes, or 10 nM RU486 was determined by quantifying MMTV RNA FISH intensity (solid bars) and by quantitative real-time PCR of mRNA (numbers on the graph are mean values ± SEM). (C) GFP-GR, GFP-N525, and GFP-407C localized to the MMTV promoter array as determined by combined RNA FISH and immunofluorescence microscopy.