Fig. 4

NFAT1 is stabilized in sunitinib-resistant RCC cells via hyperactivation of the PI3K/AKT/GSK-3β signaling pathway. A Relative mRNA expression level of NFAT1 in pre-treatment, response type and escape type RCC samples. The P values were shown as indicated. B and C Western blot analysis of NFAT1 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 Western blot analysis of NFAT1 expression in Sunitinib sensitive and resistant 786-O and ACHN cells. GAPDH served as an internal reference. E-I Western blot analysis of NFAT1 expression in 786-O and ACHN cells. GAPDH served as an internal reference. J Amino acid sequence of NFAT1, in which found a consensus binding motif of GSK-3β. K co-immunoprecipitation assay to show the interaction between NFAT1 and GSK-3β. L-O Western blot analysis of NFAT1 expression in 786-O and ACHN cells. GAPDH served as an internal reference. P and Q. Western blot analysis in 786-O cells transfected with sh-GSK-3βs (P) or GSK-3β inhibitors (Q) for 48 h. Cells were treated with MG132 (10uM) for 8 h before harvested. GAPDH served as an internal reference