Fig. 2

TGFβ1 induces p38 MAPK-dependent phosphorylation of GR Ser134. a Representative Western blot analysis of phosphorylated GR (pS134-GR), total GR, phosphorylated p38 MAPK (p-p38 MAPK), and total p38 MAPK protein levels in MDA-MB-231 cells treated with either vehicle control or 10 ng/mL TGFβ1 for 0.5, 1, or 2 h. Representative images of at least three independent experiments is shown (Densitometry of aggregated data from multiple experiments is shown in Supplemental Fig. 2); total p38 serves as a loading control. b MDA-MB-231 cells were treated with increasing concentrations of TGFβ1 for 1 h. Western blot analysis shows pS134-GR, total GR, p-p38, and total p38 protein levels (Densitometry of aggregated data from multiple experiments is shown in Supplementary Fig. 2). c Representative Western blot analysis of pS134-GR, total GR, p-p38, and total p38 in MDA-MB-231 cells pretreated with either 10 μM SB203580 (p38 inhibitor) or DMSO control for 30 min followed by either vehicle control or 10 ng/mL of TGFβ1 for 1 h. Representative images of at least two independent repeats are shown (Densitometry of aggregated data from multiple experiments is shown in Supplementary Fig. 2); total p38 serves as a loading control. d Patient-derive xenograft (PDX) HCI-10 cell line and were treated with 10 ng/mL of TGFβ1 and evaluated for expression of pS134-GR, total GR and total p38 (loading control) by Western blotting. Densitometry for pS134-GR levels are shown relative to vehicle control. e Western blot analysis of pS134-GR, total GR, p-p38 MAPK, and total p38 MAPK protein levels in MDA-MB-231 cells treated with either vehicle control or HGF (50 ng/mL) for 0.5, 1, 2, 24, and 48 h. Equal amounts of protein were loaded (Densitometry of aggregated data from multiple experiments is shown in Supplementary Fig. 2); total p38 serves as a loading control