Fig. 2
Overexpression of a mutated PIK3CA alters the response of endometrotic epithelial cells to insulin. A, Western blot of 12Z cell expression of phospho-AKT (Serine 473), AKT and β-Actin following PIK3CAH1047R expression and insulin treatment. Uncropped images are available in Supplemental Fig. 2. B, Volcano plot of DGE in PIK3CAH1047R 12Z cells treated with 100 nM insulin relative to vehicle, FDR < 0.001. Significant genes red. C, Euler diagram of overlapping DEG in PIK3CAH1047R + Insulin vs. PIK3CAH1047R and Insulin vs. Control. Statistic is hypergeometric enrichment. D, Violin plot of fold-change values relative to control for Insulin, PIK3CAH1047R and PIK3CAH1047R + Insulin for overlapping genes from F which are upregulated (n = 122, left) or downregulated (n = 229, right) in Insulin vs. Control. Statistic is paired t-test. E-F, GOBP enrichment among genes upregulated (E) or downregulated (F) in PIK3CAH1047R + Insulin vs. PIK3CAH1047R. G, GSEA plot of enrichment of Insulin vs. Control datasets for Hallmark UPR. H, Fold-change values for Hallmark UPR genes following insulin treatment in control or PIK3CAH1047R cells. Statistic is paired t-test. I, Normalized Enrichment Scores (NES) from GSEA for GOBP, Insulin vs. Control (x-axis) or PIK3CAH1047R + Insulin vs. PIK3CAH1047R (y-axis). Pathways with an absolute difference > 2 are red. Pathways related to ER are listed which are upregulated in PIK3CAH1047R + Insulin vs. PIK3CAH1047R (top) or for Insulin vs. Control (bottom). J, Tunicamycin dose response in Insulin, PIK3CAH1047R, PIK3CAH1047R + Insulin or control cells (left) and IC50 values (right). Statistic is unpaired t-test. *p < 0.05, **p < 0.01, ***p < 0.001. Western blots are representative of n = 2 independent experiments