Fig. 4

ITGB5 promotes HCC tumorigenesis via regulating β-catenin. a The increase of b-catenin/TCF/LEF-mediated transcription by ITGB5 in MHCC-97 L and Huh-7 cells with or without ITGB5 overexpression was monitored. The luciferase activities of the reporter TOP-FLASH or non-responsive control reporter FOP-FLASH were measured. The transcriptional activity was quantified via measuring the relative firefly luciferase activity normalized to the activity of Renilla luciferase. Data represent the mean ± SD of three independent experiments. ***p < 0.001 vs. control. b The decrease of b-catenin/TCF/LEF-mediated transcription by ITGB5 in MHCC-97 L and Huh-7 cells was monitored. The luciferase activities of the reporter TOP-FLASH or non-responsive control reporter FOP-FLASH were measured. The transcriptional activity was quantified via measuring the relative firefly luciferase activity normalized to the activity of Renilla luciferase. Data represent the mean ± SD of three independent experiments. **p < 0.01 and ***p < 0.001vs. control. c–d The protein and mRNA levels of Cyclin D1 and c-Myc were measured in MHCC-97 L and Huh-7 cells with or without ITGB5 overexpression. Data represent the mean ± SD of three independent experiments. **p < 0.01 and ***p < 0.001vs. control. e–f The protein and mRNA levels of Cyclin D1 and c-Myc were measured in MHCC-97 L and Huh-7 cells with or without ITGB5 knockdown. Data represent the mean ± SD of three independent experiments. **p < 0.01 and ***p < 0.001vs. control. g ITGB5 was knocked down in MHCC-97 L cells with or without β-catenin overexpression. The protein levels of ITGB5 and β-catenin were analysed by Western blotting. h–k cell proliferation was examined by colony formation assay(h and i). Cell migration was detected by transwell assay (j–k). Data represent the mean ± SD of three independent experiments. **p < 0.01 vs. control. l–m Representative images of tumor from the experimental groups, as indicated and graphic representation of tumor weight