Fig. 4

USP40 interacts with Claudin1 and regulates Claudin1 expression. A–B Western blotting was performed to determine the impact of knockdown or overexpression of USP40 on the expression of Claudin1 protein levels. C–D RT-qPCR was utilized to assess the influence of knockdown or overexpression of USP40 on Claudin1 mRNA levels. E Using USP40 and Claudin1 antibodies, the cellular location of USP40 (red) and Claudin1 (green) in HCC cells was examined by immunofluorescent staining and photographed by laser confocal microscopy. Scale bar, 50 μm. F HA-USP40 and Claudin1 plasmids were co-transfected into Huh7 and MHCC97H cells, and Co-IP was employed to determine the interactions among USP40 and Claudin1. G Representative IHC staining pictures of USP40 and Claudin1 were displayed. Scale bar, 200 μm. H The correlation between USP40 and Claudin1 expression in HCC tissues was determined by Pearson’s correlation (n = 20). ns: not significant. Data were displayed with typical pictures of three independent experiments. ***P < 0.001