Fig. 5From: Thrombospondin 4/integrin α2/HSF1 axis promotes proliferation and cancer stem-like traits of gallbladder cancer by enhancing reciprocal crosstalk between cancer-associated fibroblasts and tumor cellsHSF1 activation mediates the effects of the TSP-4/integrin α2 axis on EMT and CSC-like features in GBC cells. a The protein expression levels of p-HSF1 (sc326), HSF1 and its downstream targets after rh-TSP-4, rh-TSP-4 + anti-α2 or anti-α2 treatments were determined by western blot analysis in GBC-SD and NOZ cells. b Representative images of the Matrigel invasion assay after rh-TSP-4, si-HSF1or rh-TSP-4 + si-HSF1 treatments in GBC-SD and NOZ cells. Scale bars = 50 μm. n = three independent experiments, **P < 0.01 by ANOVA versus si-control group. c Representative images of the tumorsphere formation assay after rh-TSP-4, si-HSF1or rh-TSP-4 + si-HSF1 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 < 0.01by ANOVA versus si-control group. d The expression of EMT and CSC markers (E-cadherin, Vimentin, CD44, Nanog, Oct4 and Sox2) after rh-TSP-4, si-HSF1or rh-TSP-4 + si-HSF1 treatments were evaluated by western blotting. e The ALDH+ cells populations after rh-TSP-4, si-HSF1or rh-TSP-4 + si-HSF1 treatments were detected by Flow cytometric analysis. n = three independent experiments, **P < 0.01by ANOVA versus si-control groupBack to article page