Fig. 5
From: NEK8 regulates colorectal cancer progression via phosphorylating MYC
NEK8 phosphorylated c-MYC at serine 405. A Analysis of the potential binding of NEK8 and c-MYC using the GENEMANIA database. B, C Co-IP assays to investigate the potential interaction of NEK8 and c-MYC in SW48 (B) or Lovo (C) cells. D MG132 to inhibit endogenous c-MYC degradation; co-IP and western blot to investigate the poly-ubiquitin level of c-MYC in SW48 and Lovo stably expressing Vector or NEK8. E MG132 to inhibit endogenous c-MYC degradation; co-IP and western blot to investigate the poly-ubiquitin level of c-MYC in SW48 and Lovo expressing Vector, NEK8 WT, or NEK8 enzyme deficient mutants (NEK8-FL-KD and NEK8-FL-jck). F MG132 to inhibit endogenous c-MYC degradation; co-IP and western blot to investigate the poly-ubiquitin level of c-MYC in SW48 and Lovo stably expressing sgNC or sgNEK8. G Separately transfecting Flag-c-MYC segmented plasmid into HEK293T cells; co-IP and western blot to investigate the potential binding between c-MYC and NEK8. H Co-transfecting Vector or HA-NEK8 and Flag-c-MYC mutants into HEK293T cells; co-IP and western blot to investigate the phosphorylation level of c-MYC. I Transfecting Vector or HA-NEK8 into cancer cells stably expression c-MYC WT or c-MYC S405A, MG132 (5 uM) to inhibit endogenous c-MYC degradation; Anti-P–c-MYCS405 antibody to detect the S405 phosphorylation level of c-MYC. All western blots were conducted three times, and similar results were found