Fig. 2
Down-regulated CLEC3B in exosomes promotes migration, invasion and EMT of HCC cells. a Representative images and statistical data of relative migratory number of HCC cells with CLEC3B overexpression (Huh-7, 3B, P = 0.0086) or CLEC3B knockdown (Bel-7402, 3B-KD, P = 0.0542) and treated with exosomes from HCC cells with CLEC3B overexpression (EXO-3B, P = 0.0401) or CLEC3B knockdowned (Exo-3B-KD, P = 0.0140) in transwell assays. b Representative images and statistical data of relative invasive number of HCC cells with 3B (P = 0.0185) or 3B-KD (P = 0.0198) and treated with EXO-3B (P = 0.0003) or Exo-3B-KD (P = 0.0140) in invasive assays. c Representative images of cellular location of CLEC3B in exosomes. Green, CD63; red, Flag-CLEC3B; blue, DAPI. d The protein level of CLEC3B in exosomes secreted by HCC cells. e The mRNA expression of E-cad, N-cad and ZO-1 in HCC cells with CLEC3B overexpression (3B; E-cad, P = 0.0432; N-cad, P < 0.0001; ZO-1, P = 0.0485) or knockdown (3B-KD; E-cad, P = 0.0017; N-cad, P = 0.0461; ZO-1, P = 0.1715). The mRNA expression of E-cad (Exo-3B, P = 0.0266; Exo-3B-KD, P = 0.0047), Slug (Exo-3B, P = 0.0069; Exo-3B-KD, P = 0.0434) and ZO-1 (Exo-3B, P < 0.0001; Exo-3B-KD, P = 0.0004) in HCC cells treated with TDEs. f The protein expression of EMT relative molecules in HCC cells with CLEC3B overexpression or CLEC3B knockdown and E-cad, Slug and ZO-1 in tumor cells treated with Exo-3B and Exo-3B-KD. g Analysis of correlation of CLEC3B with E-cad (R = 0.372, P < 0.001), N-cad (R = − 0.216, P = 0.019) or ZO-1 (R = 0.180, P = 0.029) with IHC staining in hepatocellular carcinoma. h In vivo effect of CLEC3B in orthotopic transplantation model. The representative images of tumor-bearing mice (n = 6 each group) were taken at 35 days after tumor transplantation. Statistical data of tumor intensity (day 0, P = 0.9237; day 35, P = 0.0407) and metastatic loci number (P = 0.0088) were shown. 3B, CLEC3Bhigh; 3B-KD, CLEC3Blow; Exo-3B, exosomes from CLEC3Bhigh HCC cells; Exo-3B-KD, exosomes from CLEC3Blow HCC cells. *. P < 0.05; **, P < 0.01; ***, P < 0.001; n.s., not significant