Fig. 1
From: Cryptic glucocorticoid receptor-binding sites pervade genomic NF-κB response elements
DNA-binding domain mutations alter recruitment of GR to native target genes. a–c HEK293T cells were transfected with the indicated expression plasmids. The next day, the cells were steroid-deprived and treated with or without 1 μg/ml doxycycline (Dox) for 24 h. a Whole cell lysates were analyzed by western blotting using anti-HA and anti-ERK1/2 antibodies. Uncropped blots are shown in Supplementary Figure 7. b, c HEK293T cells stimulated for 1 h with 100 nM dexamethasone (Dex) alone or in combination with 10 ng/ml TNF-α as indicated and analyzed by ChIP assay using anti-HA antibody. Shown as fold enrichment relative to vehicle (mean + SEM, n = 3) *Dunnett’s multiple comparisons test, padj < 0.05 relative to GR-WT transfectants. d HEK293T cells were transfected with p65/RelA, rtTA, and the inducible HA-GR- WT or -mutant expression plasmids. The next day, the cells were steroid-deprived and treated with or without 1 μg/ml Dox for 24 h and then stimulated with 10 ng/ml TNF-α for 1 h. Whole cell lysates were analyzed by IP using anti-HA antibody or normal (pre-immune) rabbit IgG, followed by western blotting using anti-p65 antibody
