Extended Data Fig. 5: Nuclear IDO1 is degraded upon low ROS.
From: IDO1 regulating ROS rhythm reveals glycogenolysis/PPP as a cancer treatment target
(a) The expression of cytosolic or nuclear IDO1 in MCF-7 cells treated with 50 or 100 μM H2O2, or combined with 5 mM NAC was analyzed by Western blot. (b) The expression of cytosolic or nuclear IDO1 in U2OS cells treated with 100 μM H2O2 or combined with pre-treated with 5 mM NAC for 12 h was analyzed by Western blot. (c) KEAP1, TRIM21 and TRIM25 in immuno-precipitations of IDO1 were analyzed by Western blot. (d, e) The expression of nuclear IDO1 in TRIM21 KO MCF-7 (d) or TRIM25 KO MCF-7 cells (e) were analyzed by Western blot. (f) The ubiquitination of nuclear IDO1 in CUL3 KO MCF-7 cells was analyzed by Western blot. (g) The expression of nuclear IDO1 in MCF-7 cells transfected with IDO151-54 mutant plasmid was analyzed by Western blot. (h) The ubiquitination of nuclear IDO1, the binding of IDO1 and KEAP1 in synchronized MCF-7 cells at different time points was analyzed by Western blot. (i) The ubiquitination of cytosolic IDO1 in synchronized MCF-7 cells at different time points was analyzed by Western blot. For a-i, n = 3 biological independent experiments.
