Fig. 7

Overexpression of HSF1 in GBC induced the recruitment of CAFs through TGFβ signaling. a The double IF staining in human GBC tissue displayed that HSF1 was principally expressed in nuclear of gallbladder cancer cells, while α-SMA positive stroma was surrounded in HSF1 positive GBC cells. b Representative images of subcutaneous xenografts in nude mice implanted with GBC-SD cells with Vector or overexpression of HSF1 group (n = 6). c, d Xenografts weight (mg) and tumor sizes were monitored and undergone quantification analysis. n = 6, **P < 0.01 by Student’s t-test for tumor weight; **P < 0.01 by repeated-measures ANOVA for tumor sizes. e IHC analyses verified that α-SMA expression levels were significantly increased in xenograft tumors from nude mice subcutaneous implantation models of GBC-SD cells expressing exogenous HSF1. Magnification is × 400, the scale bar represents 20 μm. f Xenograft tissues arising from HSF1 overexpression group (n = 6) and control group (n = 6) were subjected to immunoblotting for HSF1, α-SMA, p-SMAD3 and t-SMAD3 protein expression, respectively. n = 6, **P < 0.01 by Student’s t-test. g The expression patterns in mRNA levels of the selected chemokines, inflammation-related genes and TGFβ after manipulation of HSF1 in GBC cells. n = three independent experiments, *P < 0.05 or **P < 0.01 by Student’s t-test. h The alterations of TGFβ1 and TGFβ2 expression levels were assessed by Western blot in GBC-SD and NOZ cells after manipulations of HSF1. β-Actin was used as an internal control. n = three independent experiments, **P < 0.01 by Student’s t-test. i The migration capacity of PTFs in response to CM-Vector, CM-OE-HSF1, CM-Vector+TGFβ1 or CM-OE-HSF1 + anti-TGFβ treatment was detected by Transwell-migration assay. Scale bars = 50 μm. n = three independent experiments, *P < 0.05 or **P < 0.01 by ANOVA