Fig. 5

RNF7 interacts with and induces the ubiquitination of SOCS1. A Cell lysates were subjected to immunoprecipitation with control IgG, anti-RNF7, or anti-SOCS1 antibodies. Then, the immunoprecipitants were blotted with the indicated antibodies. B Immunofluorescence assay of RNF7 and SOCS1 in Caki-1 and ACHN cells. Scale bar: 20 µm. C ACHN cells were transduced with RNF7-overexpressing vector and subjected to MG132 (10 µM) treatment. The mRNA and protein expression levels of RNF7 and SOCS1 were determined by quantitative RT-PCR (top) and Western blot (bottom). D RNF7-overexpressing ACHN cells were immunoprecipitated with SOCS1 or IgG antibodies, and ubiquitination was evaluated by Western blot using anti-ubiquitin antibodies. E Immunohistochemistry staining for RNF7 and SOCS1 proteins in ccRCC tissues from patients in a hospital cohort. Scale bar: 50 µm. F Correlation study of RNF7 and SOCS1 in ccRCC tissues from patients in a hospital cohort. Experiments performed in triplicate; data expressed as mean ± SD (n = 3). ***P < 0.001 compared with vector