Fig. 5: S415 glycosylation of c-Myc regulates PDK2 transcription and metabolism.
From: The OGT–c-Myc–PDK2 axis rewires the TCA cycle and promotes colorectal tumor growth
A Generation of stable HT-29 cells with c-Myc knockdown and reconstituted expression of shRNA-resistant WT or S415A c-Myc. The c-Myc knockdown efficiency and re-expression were examined using immunoblotting. B Quantitative PCR analysis was performed to detect the expression of the PDK2 in HT-29 cells with c-Myc knockdown and reconstituted expression of shRNA-resistant WT and S415A c-Myc in the presence or absence of OGT overexpression. n = 3; Data are presented as means ± SD. P values were determined by unpaired two-tailed Student’s t tests. C Dual-luciferase reporter assay showing the effects of c-Myc WT and S415A on relative PDK2-promoter activity in the presence or absence of TMG in the 293 T cells. n = 3; Data are presented as means ± SD. P values were determined by unpaired two-tailed Student’s t tests. D Immunoblotting of c-Myc and PDK2 expression in stable HT-29 cells ectopically expressing Flag-tagged WT or S415A c-Myc with PDK2 overexpression, with a simultaneous depletion of the endogenous c-Myc. E ECAR and OCR in HT-29 cells stably expressing scramble shRNA, c-Myc-targeting shRNA and shRNA-resistant WT, S415A c-Myc or S415A c-Myc with PDK2 overexpression. n = 3; Data are presented as means ± SD. P values were determined by unpaired two-tailed Student’s t tests. Comparison of glucose uptake (F), ATP(G), lactate (H), GSH/GSSG (I), ROS (J) in HT-29 cells reconstituted with WT, S415A c-Myc or S415A c-Myc with PDK2 overexpression. n = 3; Data are presented as means ± SD. P values were determined by unpaired two-tailed Student’s t tests.
