ZDHHC2 Antibody - #DF4688

Figure 1. ZDHHC2-mediated S-palmitoylation plays a key role in modulating the sensitivity of ccRCC to sunitinib. A, 786-O cells were transfected with indicated constructs for 72 hours. These cells were harvested and treated with a serial dose of sunitinib for 24 hours. The CCK-8 assay was applied to measure the IC50 values of sunitinib in each group. Data presents as mean ± SEM with three replicates. B, Expression levels of ZDHHCs during sunitinib pretreatment (n = 4), response (n = 4), and resistance (escape; n = 4) phases. The heatmap shows the log-fold change (LogFC) of escape versus pretreatment and log-fold change of escape versus response. C, The protein level of ZDHHC2 from patients with RCC with (n = 6) or without (n = 6) sunitinib resistance was examined by Western blotting analysis, and the protein level of ZDHHC2 was quantified by ImageJ software. P values as indicated. D, The protein level of ZDHHC2 from 786-O cells with or without sunitinib resistance (786-O R) was examined by Western blotting analysis. Three replicates were performed. E–G, 786-O cells were transfected with indicated constructs for 48 hours. After 24-hour puromycin selection, cells were treated with or without sunitinib (2 μmol/L) for another 24 hours. Cells were collected for Western blot analysis (E), caspase-3 activity assay (F), and Annexin V-FITC/7-AAD assay (G). Data presented as mean ± SEM with three replicates. H, 786-O, A498, and 786-O R cells were transfected with indicated constructs for 72 hours. After puromycin selection, these cells were treated with a serial dose of sunitinib for 24 hours and subjected to CCK-8 assay. The IC50 values of sunitinib in each group are indicated. I–K, 786-O cells were transfected with indicated constructs for 72 hours. After puromycin selection, these cells were subcutaneously injected into the nude mice. These mice were treated with or without sunitinib (oral administration, 25 mg/kg, once a day for 8 days). Tumor image (I); tumor mass (J); tumor growth curve (K). Data presented as mean ± SEM with five replicates. L and M, 786-O cells were transfected with indicated constructs for 24 hours. Cells were treated with or without sunitinib (2 μmol/L) for another 24 hours. Cells were collected for Western blot analysis (L) and caspase-3 activity assay (M). Data presented as mean ± SEM with three replicates. N, 786-O and A498 cells were transfected with indicated constructs for 24 hours. These cells were treated with a serial dose of sunitinib for 24 hours and subjected to CCK-8 assay. The IC50 values of sunitinib in each group are indicated. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. EV, empty vector.