Figure 2.

Induction of BAG3 upon BCSC-enrichment culture. (a and b) Breast cancer cells were cultured under traditional adhesive (Ad) or mammosphere-forming (Ma) culture, cancer stem cell markers SOX2 (a) and NANOG (b) expression was analyzed by real-time PCR. (c and d) Breast cancer cells were cultured as above, BAG3 mRNA (c) and protein (d) expression was analyzed by real-time PCR and western blot, respectively. (e) Breast cancer cells were infected with lentivirus containing empty (Con) or gRNA directed BAG3 (KD); BAG3 expression was analyzed by western blot. (f) Control (Con) or BAG3 knockdown (KD) breast cancer cells were cultured under mammosphere-forming condition. Mammospheres were photographed by phase-contrast microscopy and the number of mammospheres was counted, and was represented as the mean±S.E.M. from three independent experiments. NS, not significant; *P<0.01