Fig. 5

FBW7 promotes NFAT1 degradation in RCC cells. A Amino acid sequence of NFAT1, in which found a consensus binding motif of FBW7. B Relative mRNA expression level of FBW7 in pre-treatment, response type and escape type RCC samples. The P values were shown as indicated. The mean of each group was compared with the mean of every other group. C Western blot analysis of FBW7 expression in Sunitinib sensitive and resistance RCC patients. GAPDH served as an internal reference. ***, P < 0.001. The difference was compared between sunitinib sensitive group and sunitinib resistance group. D Scatter diagram to show the correlation between the protein expression level of NFAT1 and FBW7 in renal cancer patients. P values as indicated. E co-immunoprecipitation assay to show the interaction between NFAT1 and FBW7 in 786-O and ACHN cells. F-K Western blot analysis of NFAT1 expression in 786-O and ACHN cells with different treatment, which was indicated in the figure labels. GAPDH served as an internal reference. L-M Western blot analysis in 786-O cells transfected with shFBW7s (L) or FBW7 plasmids (M) for 48 h. Cells were treated with MG132 (10 uM) for 8 h before harvested. GAPDH served as an internal reference. N 786-O cells were infected with indicated constructs for 48 h. Then, cells were treated with or without MK2206 (10 uM) for another 24 h and subjected to Western blot analysis. O 786-O cells were infected with indicated constructs for 48 h. Cells were subjected to Western blot analysis. P 786-O cells were transfected with indicated constructs for 24 h. Cells were subjected to Western blot analysis