Fig. 3

RNF31 restrains TNBC cell progression via Hippo/YAP axis. A Western blot detecting of YAP and RNF31 expression in MDAMB231 cell exposed to siControl or siRNF31 or siRNF31 + siYAP for 48 h, respectively. B RT–qPCR results of CTGF and CYR61 mRNA expression in MDAMB231 cell line exposed to indicated treatment. C-D Transwell assay (left panel) of MDAMB231 cells transfected with indicated treatment. Right panel shows quantification of transwell assay results. Scale bar 100 μm. E-F Representative images (left panel) of EdU assays in MDAMB231 cell transfected with indicated treatment. EdU-positive cells, red; cell nuclei, blue. Right panel shows quantification of Edu results. Scale bar 100 μm. G-H FACS analysis (left panel) was performed on the MDAMB231 cell to detect the proportion of apoptotic cells. I-J Wound healing assay (left panel) of MDAMB231 cell migration capability. K-L FACS analysis (left panel) was performed on the MDAMB231 cell to determine the proportion of CD44 + CD24-cells. M Representative image of tumor derived from BALB/c nude mice injected with indicated stably transfected MDAMB231 cells. N-O The tumor volume (N) and weight (O) in BALB/c nude mice subcutaneously inoculated with indicated stably cells. P Western blot detecting of YAP and RNF31 expression in tumor tissues in BALB/c nude mice injected with indicated stably cells. Q Representative image of In vivo lung metastasis of indicated MDAMB231 cells. R-S Representative HE stains pictures of lung sections of indicated MDAMB231 cells. Arrows indicate metastatic nodules (R). Quantification of metastatic lung nodules (S). The results are representative of 3 independent experiments in panel A-L. The results are representative of 6 independent experiments in panel M-Q. The results are representative of 4 independent experiments in panel Q-S. β-actin was engineered to the internal reference for Western blot. The data are presented as mean ± SDs. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t test)