Extended Data Fig. 1: ROS rhythm is regulated by IDO1 in human tumor cells.

(a) The intensity of HyPerRed fluorescence of synchronized SKOV3 and HCT116 cells were detected by flow cytometry for 96 h. Time 0 was the first time point after synchronization, and cells were collected according to different time points for detection. (b, c) The intensity of HyPerRed fluorescence and GFP fluorescence of an individual synchronized MCF-7 or U2OS cells were detected using time-lapse microscopy for 48 h. Scale bar, 10 μm (b). The intensity of HyPerRed fluorescence and GFP fluorescence were recorded(c). (d) The intensity of HyPerRed fluorescence and SypHer fluorescence in synchronized MCF-7 cells at different time points were recorded. HyPerRed, Ex/Em=575/605 nm; SypHer, Ex/Em=445/510 nm. (e, g, i) The intensity of HyPerRed fluorescence of synchronized MCF-7 and U2OS cells pre-treated with 20 μM CLK8 for 12 h were detected by flow cytometry for 48 h.(e), synchronized BMAL1 knockout MCF-7 and U2OS cells (g), synchronized MCF-7 and U2OS cells transfected with si-NC or si-PRDX1 (i). (f) The expression of BMAL1 in MCF-7 and U2OS cells targeted BMAL1 by CRISPR/Cas9 was analyzed by Western blot. (h) The expression of PRDX1 in MCF-7 and U2OS cells transfected with si-NC, si-PRDX1 were analyzed by Western blot. For a-i, n = 3 biological independent experiments. JTK_CYCLE analysis was used to determine rhythmicity (a), and P < 0.05 was considered to be rhythmic.

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