Fig. 5: WTAP activates AKT expression via an m6A-dependent manner to promote enzalutamide resistance.

A Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showing altered pathways between C4-2B and C4-2B-ENZR cells. B Immunoblots of indicated proteins in C4-2B parental and C4-2B-ENZR cells. C4-2B-ENZR cells were transfected with siRNAs targeting WTAP. The relative AKT, p-AKT, and WTAP quantification of the Western blotting was measured by using ImageJ software. *P < 0.05. C 22Rv1 cells were transfected with siCtrl or WTAP siRNA. Western blotting was performed to detect AKT expression. The relative AKT and WTAP quantification of the Western blotting was measured by using ImageJ software. *P < 0.05. D, E C4–2B-ENZR and 22Rv1 cells were treated with STM2457 at the indicated concentrations for 48 h. Western blotting was used to detect the level of AKT. The relative AKT quantification of the Western blotting was measured by using ImageJ software. *P < 0.05. F AKT mRNA stability was measured in C4-2B and C4-2B-ENZR cells. G MeRIP-qPCR analysis was used to validate m6A levels of AKT pre-mRNA in C4-2B and C4-2B-ENZR cells. H MeRIP-qPCR analysis was used to validate m6A levels of AKT pre-mRNA in C4-2B shCtrl and shWTAP cells. I–K Cell viability was measured by Cell Counting Kit-8 (CCK8) assay in C4-2B and C4-2B-ENZR cells under AKT inhibitor or enzalutamide treatment, or USP7 inhibitor. Cells were transiently transfected with the indicated WTAP siRNA or Flag-WTAP plasmid before treatment. *P < 0.05; **P < 0.01, ***P < 0.001.