Fig. 4: Regulation of LRRC58–CDO1 by cellular cysteine abundance.
From: Covariation MS uncovers a protein that controls cysteine catabolism
a, Post-translational stability of CDO1 reporter over time following switch from standard medium (0.1 mM cystine) to medium without cystine. Data are normalized to standard medium. t = 0, n = 10 cell replicates; other time points, n = 5 cell replicates. Statistical comparison is to t = 0. b, Post-translational stability of CDO1 following exposure to media with indicated levels of d-cysteine and l-cysteine for 24 h. Data are normalized to cells maintained in medium without cystine. No-cystine control, n = 8; other treatments, n = 4. Statistical comparison is between d-cysteine and l-cysteine. c, Reporter cells were cystine-depleted for 24 h, then changed back to normal 0.1 mM cystine medium for the indicated length of time. GFP/mCherry ratio is normalized to cells that were maintained in 0.1 mM cystine. n = 4 cell replicates. Statistical comparison is to t = 0. d, Concentration-dependent effect of cysteine on post-translational stability of CDO1 in LRRC58KD or scr cells. n = 4 cell replicates. Statistical comparison is between LRRC58KD and scr cells at each cysteine concentration. e, AlphaFold predicted structure of the interaction between LRRC58 and CDO1, with interface residues labelled. f, GFP/mCherry ratio in LRRC58KD and scr Hep G2 cells expressing CDO1 mutant reporters. n = 4 cell replicates. g, GFP/mCherry ratio in Hep G2 cells expressing CDO1 mutant reporters treated with 0.1 mM cystine or cystine-restricted. n = 4 cell replicates. Two-tailed Student’s t-test for pairwise comparisons. Data are mean ± s.d.
