Fig. 5

YAP1 is a direct target of miR-375. a Schematic illustration showing complementarity to the miR-375 seed sequence in the 3′-UTR of YAP1. Lowercase letters indicate mutated nucleotides. b YAP1 expression was higher in human OS than in chondroma tissue. Data represent the mean ± SD (n = 12). c YAP, c-Myc, and Birc5 expression was higher in human OS than in chondroma tissue. Representative images are shown (Scale bars = 100 μm). d 293T cells were co-transfected with pre-miR-375 and a luciferase reporter construct containing wild-type (WT) or mutated YAP1 3′-UTRs. Data represents the mean ± SD (n = 3). * P < 0.05. e–f miR-375 overexpression reduced YAP1, c-Myc, and Birc5 (e, f) protein and (g) mRNA levels while miR-375 inhibition increased YAP1, c-Myc, and Birc5 (e, f) protein and (g) mRNA levels. Cells were transfected with NC or miR-375 mimics/inhibitor, and mRNA or protein levels were evaluated. Protein levels were evaluated by western blot and immunofluorescence, mRNA levels were evaluated by qRT-PCR. Data represent the mean ± SD (n = 3). * P < 0.05. Scale bars = 50 μm. h Overexpression of YAP1 inhibits anoikis in osteosarcoma cells. HOS and 143B stable cells were subjected to suspension culture for 48 h. Anoikis rates were determined by Annexin V-FITC/PI staining. Data represent the mean ± SD (n = 3). * P < 0.05