Fig. 4: The Y537S mutant ER regulated key mediators of the RON/PI3K pathway.

a ER ChIP-Seq track for GAB2 gene body in MCF-7 ESR1 Y537S cells. b ER ChIP-qPCR in MCF-7 ESR1 WT and MCF-7 ESR1 Y537S cells at the binding site highlighted in a. Graphs represent mean + standard deviation (N = 3 replicates). Student’s t-test was performed for statistical analysis. c MCF-7 cells were cultured in 10% FBS supplemented media and treated with 1 µM Tam and 1 µM Ful for 48 h. qRT-PCR analysis of GAB2 was performed. Graphs represent mean + standard error of the mean (N = 3 replicates). Two-way ANOVA was performed for statistical analysis. d MCF-7 cells were cultured in 10% FBS supplemented media and transfected with scrambled (Scr) siRNA and GAB2 siRNA. Immunoblot analysis of phosphorylated and total AKT and p44/42 MAPK was performed 48 h post transfection. e T47D cells were cultured in 10% FBS supplemented media and transfected with GAB2 siRNA. Immunoblot analysis of phosphorylated and total AKT and p44/42 MAPK was performed 48 h post transfection. P < 0.05 was considered statistically significant for all tests. (*p < 0.05, **p < 0.01, ***p < 0.001).