Fig. 6.

YAP mediates the tumor-suppressive function of miR-375 in OS cells in vitro. a The expression of YAP1, c-Myc, and Birc5 in HOS and 143B cells was detected by immunofluorescence analysis. Cells were co-transfected with miR-375 and YAP1 plasmids or control vector. Data represents the mean ± SD (n = 3). Scale bars = 50 μm. b Downregulation of YAP1 and miR-375 induced more anoikis in osteosarcoma cells compared with miR-375 knockdown alone. HOS and 143B stable cells were subjected to suspension culture for 48 h. Anoikis rates were determined by Annexin V-FITC/PI staining and FACS. Data represents the mean ± SD (n = 3). * P < 0.05. c Proliferation of OS cells, transfected with pre-miR-375 or control vector with or without YAP1, was evaluated by the CCK-8 assay. Data represents the mean ± SD from three independent experiments. d YAP1 overexpression stimulated the growth of miR-375 overexpression cells, as determined by the colony formation assays (details are shown in the insets). Data represent the mean ± SD (n = 3). * P < 0.05. e Effects of miR-375 overexpression on cell migration and invasion were abrogated by YAP1 overexpression. Migration and invasion of OS cells co-transfected with pre-miR-375 and YAP or control shRNA were evaluated with the Matrigel and Transwell invasion assays, respectively. Scale bars = 50 μm. f The effect of miR-375 and YAP overexpression on cell migration was evaluated by wound healing assay in HOS and 143B cells, respectively. Scale bars = 200 μm