Fig. 5. Inhibition of endogenous GP73 blocks trafficking of intracellular MMP-7.

a The levels of GP73 and MMP-7 in indicated normal liver and HCC cell lines were determined by immunoblotting analysis. The correlation between levels of GP73 and MMP-7 in indicated cells was represented using linear correlation. b Immunofluorescence staining of GP73 (red) and MMP-7 (green) in primary tumor (n = 90) and adjacent liver (n = 90) tissues of HCC patients. Images are labeled by stage (1, 2, 3) and tissue type (C = primary tumor, N = adjacent liver, scale bar: 25 μm). The expression of specific proteins was measured using ImageJ and represented as the average optical density (AOD). The scatter plot indicates the relative expression of GP73 and MMP-7. The correlation between GP73 and MMP-7 in carcinoma tissues was represented using linear correlation. c Immunoblotting analysis and immunofluorescence staining (red: GP73, green: MMP-7; scale bar: 10 μm) of GP73 and MMP-7 after MHCC-97H cells were transfected with siGP73 (20 nM) for 0, 6, 12, 24, 48, and 72 h. Cell culture media were collected for ELISA before the cells were harvested. d Immunoblotting analysis and immunofluorescence staining (red: GP73, green: MMP-7; scale bar: 10 μm) of GP73 and MMP-7 after MHCC-97H cells were treated with BFA (2.5 μg/ml) for 0, 0, 5, 1, 2, 6, and 12 h. Cell culture media was collected for ELISA before cells were harvested. Data in a, b, c and d are the mean ± s.e.m. A two-tailed Student’s t-test was used for statistical analysis