OTUD1 sensitizes ccRCC cells to the TKIs via PTEN. A, 786-O and ACHN cells were transfected with indicated constructs for 48 h. Cells were treated a serial concentration of Sunitinib for measuring the IC50 values of Sunitinib. B, 786-O and ACHN cells were infected with indicated constructs for 72 h. Cells were treated a serial concentration of Sunitinib for measuring the IC50 values of Sunitinib. C and D, 786-O and ACHN cells were transfected with indicated constructs for 48 h. Cells were treated with 1μM Sunitinib for another 24 h and cells were harvested for Western blot (C) and RT-qPCR analysis (D). Data presents as mean ± SD with three replicates. Ns, not significant; *, P < 0,05; **, P < 0.01; ***, P < 0.001. E and F, 786-O and ACHN cells were infected with indicated constructs for 72 h. Cells were treated with 1μM Sunitinib for another 24 h and cells were harvested for Western blot (E) and RT-qPCR analysis (F). Data presents as mean ± SD with three replicates. Ns, not significant; *, P < 0,05; **, P < 0.01; ***, P < 0.001. G, 786-O and ACHN cells were infected with indicated constructs for 72 h. Then, these cells were pretreated with or without IKK-16 (500 nM) and treated a serial concentration of Sunitinib for measuring the IC50 values of Sunitinib. H, 786-O and ACHN cells were infected with indicated constructs for 72 h. Then, these cells were pretreated with or without JSH-23 (10 μM) and treated a serial concentration of Sunitinib for measuring the IC50 values of Sunitinib. I, 786-O and ACHN cells were infected with indicated constructs for 72 h. Then, these cells were pretreated with or without MK2206 (10 μM) and treated a serial concentration of Sunitinib for measuring the IC50 values of Sunitinib. J, a model depicting that the downregulation of OTUD1 stabilizes PTEN to inactivate AKT and TNF/NF-kappa B pathway and be responsible for the Sunitinib resistance in ccRCC.