Fig. 6
FBW7 contributes to modulating the immune response in RCC. A and B. Western blot (A) and qRT-PCR (B) analysis of PD-L1 expression in renal cancer cells infected with shControl or shFBW7s. GAPDH served as an internal reference. Data presented as the mean ± SD of three independent experiments. ***, P < 0.001. The difference was compared between shControl group and shFBW7 group. C and D. Western blot (C) and qRT-PCR (D) analysis of PD-L1 expression in renal cancer cells infected with EV or FBW7α plasmids. GAPDH served as an internal reference. Data presented as the mean ± SD of three independent experiments. ***, P < 0.001. The mean of each group was compared with the mean of every other group. E and F. Western blot (E) and qRT-PCR (F) analysis of PD-L1 expression in renal cancer cells infected with or without shFBW7 and/or shNFAT1 #1. GAPDH served as an internal reference. The mean of each group was compared with the mean of every other group. G and H. Western blot (G) and qRT-PCR (H) analysis of PD-L1 expression in renal cancer cells infected with or without FBW7 plasmids and/or shNFAT1 #1. GAPDH served as an internal reference. The mean of each group was compared with the mean of every other group. I The correlation between interested immune cell subsets and the expression of FBW7 in renal cell carcinoma. J-K TIMER database were searched to determine the correlation between CD4+ T cells (J) and CD8+ T cells (K) infiltration in RCC. L Western blot analysis of Fbw7 and PD-L1 expression in Renca cells infected with Ev or Fbw7 plasmids. GAPDH served as an internal reference. M After 72 h of selection with puromycin, 5 × 106 Renca cells infected with Ev or Fbw7 plasmids were subcutaneously injected into the right dorsal flank of C57BL/6 mice. Mice with subcutaneous Renca tumors (n = 5/group) were treated with anti-PD-1 (200 μg) or nonspecific IgG for three times at day 1, 4, and 7. N Immunofluorescence staining analysis of the percentage of CD3+CD4+ and CD3+CD8+ T cells infiltrated in Renca tumors. Data are presented as the mean ± SD of five independent experiments (***, P < 0.001). The mean of each group was compared with the mean of every other group