Figure 2.

Loss of BGN in BCSCs leads to reduced CSC populations and tumorsphere forming ability. (A) Immunoblots showing BGN expression in MMTV-PyMT tumor cells (FF99) were stably expressing mouse BGN sgRNA and Control sgRNA (Ctrl sgRNA). Immunoblots and immunofluorescence depicting BGN expression in (B,C) ALDH⁺ BCSCs and (D,E) CD29^hi CD61⁺ BCSCs. (F) Dot plots of ALDH assay of ALDH⁺ cells in BGN sgRNA cells compared to Ctrl sgRNA. (G) Quantification of ALDH assay of ALDH⁺ BCSCs with BGN depletion. (H) FACS analysis of CD29 and CD61 marker expression in FF99 cells transduced with control and BGN sgRNA. (I) Quantification of H depicting the percentage of CD29^hi CD61⁺ BCSCs of BGN depleted cells. Representative images showing tumorsphere formation of (J) CD29^hi CD61⁺ and (L) ALDH⁺ BCSCs and their (K,M) quantification. Scale bar = 50 µm. Data shown as mean ± SEM of three or four independent experiments. Statistical significance was determined using two-tailed t test; * p ≤ 0.05 as compared to control sgRNA. Representative images of number of spheres were quantified using ImageJ.