Fig. 3

OSM signaling promotes an EMT that is independent of LOXL2 expression. a Confocal images of MCF7 cells depict a distinct loss of cell polarity and a transition from membrane localization to cytoplasmic localization of E-cadherin with 48-h OSM treatment, both hallmarks of EMT. E-cadherin (red) and nuclei (DAPI, blue). Magnification × 40 with digital zooming × 2; scale bar = 20 μm. b Immunoblot of MCF7-sh-non-target and MCF7-sh-LOXL2 cells treated with OSM for 24 and 48 h. Expression of EMT markers, E-cadherin (E-Cad), and Snail are compared between these cell lines. The absence of LOXL2 expression in MCF-7 cells had no effect on OSM-induced EMT. c Immunoblot of MDA-MB-468 breast cancer cells exposed to siCTRL and siLOXL2 for 48 h prior to OSM treatment for 24 and 48 h. Expression of EMT markers, E-Cad, and Snail are compared between siLOXL2 and siCTRL treatments. Inhibited LOXL2 expression in MDA-MB-468 cells had no effect on OSM-induced EMT. d Immunoblot of MCF7 cells treated with OSM for 24 h; post treatment cells are collected and subjected to nuclear-cytoplasmic protein fractionation. LOXL2 protein expression is not present in the nuclear fraction, only in the cytoplasmic fraction. GAPDH protein expression is used to confirm purity of cytoplasmic fraction, and Snail transcriptional factor expression is used to confirm nuclear fraction purity. (All experiments n = 3+)