Figure S4 from Tissue Stiffness and Hypoxia Modulate the Integrin-Linked Kinase ILK to Control Breast Cancer Stem-like Cells

Depleting ILK inhibits PI3K signaling in human breast cancer cells. Immunoblotting analysis for phosphorylated and total Akt in (A) MDA-MB-231-shctrl or MDA-MB-231-shILK cells and (B) MDA-MB-231 cells treated with or without LY294002 (50 µM). Transcript levels of (C) ITGB1, (D) ILK, (E) CD44, (F) Nanog, and (G) CD49f in MDA-MB-231 cells treated with or without LY294002 and cultured on soft or stiff substrata under normoxia or hypoxia. Phase contrast images and quantification of (H) secondary mammospheres and (I) colonies formed in soft agar by MDA-MB-231 cells treated with or without LY294002 under normoxia or hypoxia. Scale bars, 50 µm. Shown are mean (plus minus) s.e.m. *, P<0.05; **, P<0.01; ***, P<0.001.