Figure S3 from Tissue Stiffness and Hypoxia Modulate the Integrin-Linked Kinase ILK to Control Breast Cancer Stem-like Cells

Ectopic expression of ILK induces breast CSC development in MDA-MB-231 human breast cancer cells. (A) qRT-PCR and immunoblotting analysis for ILK and GAPDH in MDA-MB-231 cells transduced with adGFP or adILK. (B) Immunoblotting analysis for phosphorylated and total Akt in MDA-MB-231 cells transduced with adGFP or adILK. Transcript levels of (C) ITGB1, (D) CD44, (E) Nanog, and (F) CD49f in MDA-MB-231 cells transduced with adGFP or adILK and cultured on soft or stiff substrata under normoxia or hypoxia. Phase contrast images and quantification of (G) secondary mammospheres and (H) colonies formed in soft agar by MDA-MB-231 cells transduced with adGFP or adILK. Scale bars, 50 µm. Shown are mean (plus minus) s.e.m. *, P<0.05; **, P<0.01; ***, P<0.001.