Fig. 6

S405 phosphorylation of c-MYC blocked its poly-ubiquitin. A Western blots to detect the expression level of c-MYC, cyclin D1, and CDK4 in cancer cells stably expressing c-MYC WT, c-MYC S405A, or c-MYC S405E. B CCK8 assays to detect the proliferation ability of cancer cells stably expressing c-MYC WT, c-MYC S405A, or c-MYC S405E; n = 3, P(Left) = 0.0004 or 0.0002, P(Right) = 0.0071 or 0.0044. C Detecting the cellular ATP level of cancer cells stably expressing c-MYC WT, c-MYC S405A, or c-MYC S405E; n = 3, P(SW48) = 0.0001 or < 0.0001, P(Lovo) = 0.0009. D, E EdU assays to investigate the DNA replication ability of cancer cells stably expressing c-MYC WT, c-MYC S405A, or c-MYC S405E; representative images shown (D); statistical analysis shown (E); n = 3, P(SW48) = 0.002 or = 0.0008, P(Lovo) = 0.0005 or = 0.0008. F MG132 to inhibit c-MYC degradation; co-IP and western blots to detect the poly-ubiquitin level of c-MYC in SW48 cells stably expressing c-MYC WT, c-MYC S405A, or c-MYC S405E. G Cycloheximide (CHX, 50 uM) was applied to SW48 cells stably expressing c-MYC WT, c-MYC S405A, or c-MYC S405E at the indicated time; western blots to detect the degradation rate of c-MYC. H Statistical analysis for Fig. 6G; n = 3, P = 0.001 or = 0.008. All western blots were conducted three times, and similar results were found; a Student's t-test was applied for statistical analysis