Fig. 4

Integrin α2 mediates the efficacy of paracrine of TSP-4 signaling on EMT and cancer stemness of GBC cells. a Representative images of the Matrigel invasion assay after rh-TSP-4, rh-TSP-4 + anti-α2 or anti-α2 treatments in GBC-SD and NOZ cells. Scale bars = 50 μm. n = three independent experiments, **P < 0.01 or ^# P < 0.01by ANOVA versus control group. b Representative images of the tumorsphere formation assay after rh-TSP-4, rh-TSP-4 + anti-α2 or anti-α2 treatments in GBC-SD and NOZ cells. The number of tumorspheres was counted and plotted, and the percentage of tumorspheres with diameters of 50–100 μm, 100–150 μm or > 150 μm was calculated and plotted. Magnification is × 200, and scale bars = 50 μm. n = three independent experiments, **P < 0.01 or ^# P < 0.01by ANOVA versus control group. c The expression of CSC and EMT markers after rh-TSP-4, rh-TSP-4 + anti-α2 or anti-α2 treatments were evaluated by western blotting. d The ALDH+ cells populations after rh-TSP-4, rh-TSP-4 + anti-α2 or anti-α2 treatments were detected by Flow cytometric analysis. n = three independent experiments, *P < 0.05 or **P < 0.01 or ^# P < 0.01by ANOVA versus control group