Extended Data Fig. 8: IDO1-catalyzed Kyn activates G6PD to clear ROS via NADPH.
From: IDO1 regulating ROS rhythm reveals glycogenolysis/PPP as a cancer treatment target
(a) Molecular docking and Molecular dynamics simulation of G6PD and G6P with Kyn. (b) Recombinant G6PD proteins were stained with Coomassie Brilliant Blue. (c) MCF-7 cells cultured in 13C-glucose for 5 days and switched to 12C-glucose combined with 100 μM kyn treatment for 6 h. 13C-labeled R5P was detected by LC-MS/MS. n = 6 technical replicates. (d) The NADPH concentration in MCF-7 cells treated with 100 μM Kyn for 6 h was measured. n = 6 technical replicates. (e, f) The intensity of HyPerRed fluorescence of synchronized U2OS cells at different time points was detected by flow cytometry for 48 h, and Kyn concentration (e) and the NADPH/NADP+ level (f) were measured by chromatography and NADPH assay kit. (g, h) The concentration of NADPH/NADP+ level (g) and Kyn concentration (h) in synchronized IDO1 knockout MCF-7 cells were analyzed. (i) The intensity of HyPerRed fluorescence and the NADPH/NADP+ level of synchronized G6PD knockdown MCF-7 cells was analyzed. For e-i, n = 3 biological independent experiments. All error bars are mean ± SD. P values were calculated by two-tailed Student’s t test (c, d).
