Fig. 2
bCAFs exert their protective effects by paracrinely favoring MCL-1 expression. a Anti-apoptotic (BCL-2, MCL-1, BCL-xL) proteins expression levels in ZR-75-1 (top) or T-47D (bottom) cells, treated or not for 24 h with ABT-737 (1 µM) in presence of non-CM or CAFs-CM, were evaluated using western-blot analysis. b qRT-PCR of MCL-1 mRNA in ZR-75-1 cells grown in presence of non-CM or different CAFs CM for six and 12 h. Mean and SEM of three independent experiments are represented as relative quantity of mRNA normalized to the mean of RPLP0, B2M and GAPDH relative expression. c MCL-1 proteins expression levels in ZR-75-1 cells grown in presence of non-CM or CAFs-CM for 24 h and treated by cycloheximide for the indicated time, were evaluated using western-blot analysis. d ZR-75-1 cells (left) or T-47D cells (right) were treated for 48 h with ABT-737 1 µM and/or A-1210477 5 µM in presence of non-CM or Fibroblasts-CM. e ZR-75-1 cells infected by empty vector, MCL-1 or BCL-XL shRNA, were treated for 48 h with ABT-737 1 µM in presence of non-CM or fibroblasts-CM. Percentage of positive Annexin-V-FITC or –APC apoptotic cells was measured by flow cytometry (d, e). Data are means ± SEM from three independent experiments. P-value was determined by two-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: not significant
