Fig. 2
From: BCL-XL directly modulates RAS signalling to favour cancer cell stemness
BCL-XL maintains cancer initiating cell features induced by RAS activity in mammary cells. a, b Percentage of sphere forming cells in bulk population of a MCF10A KRASV12 cells or c EGF-treated MCF10A Lxsn cells after 72 h infection with the indicated lentivector sh-RNAs. As comparison, the measured lack of sphere formation of Lxsn untreated with EGF is also illustrated. 128 cells per condition were seeded in serum-free media in ultra-low adhesion plates and the resulting spheres counted after two weeks incubation. Each dot represents the percentage of cell forming sphere for one biological replicate. Mean and SEM of at least 3 (3 to 6) independent experiments are represented (two-tailed unpaired t-test). b Western blot analysis of BCL-XL expression and ERK phosphorylation in MCF10A Lxsn cells after overnight starvation and treatment with 20 ng ml−1 EGF for an additional 24 h d. Transplant experiment of MCF10A KRASV12 cells infected with sh-BCL-XL or a control vector. 5 × 105 cells were subcutaneously injected in 7 Nu/Nu mice for each group and tumour growth was monitored (Log-rank (Mantel-Cox) test). e Flow cytometry analysis of CD44 expression on MCF10A KRASV12 cells infected with sh-BCL-XL or a control vector 72 h prior staining. White histograms represent the IgG control staining; blue histograms represent the CD44 staining. Data from one representative experiment are shown
