Fig. 7

Fig. 7

ADAMTS1 is associated with p53 to modulate epidermal growth factor receptor (EGFR) promoter activity and expression in renal cell carcinoma (RCC) cells. A Luciferase reporters were transfected into Caki-1 and 786-O cells with or without ADAMTS1 manipulation. All luciferase activity was normalized to Renilla reporter activity. The normalized reporter activity in cells transfected with a control vector was considered as onefold, and the relative fold change for each construct (ADAMTS1-flag or shADAMTS1) was further normalized to that of the control. B Reporters were transfected into Caki-1 and 786-O cells and treated with conditioned media (CM) collected from ADAMTS1-overexpressing or control Caki-1 and 786-O cells. The normalized reporter activity in cells treated with CM collected from control cells was considered onefold, and the relative fold change of RCC-derived CM treatment was calculated. The values are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, compared with the control group. C Western blotting analysis of ADAMTS1 expression levels in cytoplasmic and nuclear fractions from Caki-1 and 786-O cells transfected with ADAMTS1-flag or a control vector. D The immunocomplex was precipitated from ADAMTS1-overexpressing Caki-1 cell lysates with an ADAMTS1 antibody and analyzed by western blotting to detect the associations of ADAMTS1 with indicated transcription factors (p53, WT1, and c-Jun). A normal IgG antibody was used as an immunoprecipitation (IP) control, and 10% whole-cell lysate was used as the input. E Western blot analysis of p53, ADAMTS1, p-EGFR, and EGFR levels in Caki-1 and 786-O cells after transducing ADAMTS1-flag or a control vector and treatment with the p53 inhibitor, pifithrin (PFT)-α (10 µM), or the vehicle for 24 h