Fig. 8

GPR65 is negatively correlated with the migration of OS cells. (A) Typical images of U2OS and HOS cells crossing the non-matrigel-coated membrane (transwell assay). (B) Typical images of U2OS and HOS cells at initial (0 h) and final position (24 h) (wound-healing assay). (C) U2OS and HOS cells were transfected with siNC and siGPR65 or Vector and Flag-GPR65 plasmids, followed by immunofluorescence assay to observe F-actin (green). Nuclei were stained with DAPI (blue). Scale bars: 200 μm. (D) The changes of EMT markers protein levels after GPR65 knockdown or overexpression. (E) Data of numbers of invasive cells per field. (F) Data of relative migration index. (G) Data of the green fluorescence intensity of F-actin. (H-I) qRT–PCR showed the changes in EMT markers after GPR65 knockdown or overexpression. Data are mean ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 compared with the control group