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. 2022 Apr 19;11:e76021. doi: 10.7554/eLife.76021

Figure 5. Methionine and choline supplementation alleviate dithiothreitol (DTT) toxicity.

Figure 5.

(A) Representative images of N2 and metr-1(ok521) animals after 5 days of hatching at 20°C on E. coli OP50 diet containing 10 mM DTT, 10 mM DTT supplemented with 10 mM methionine, and 10 mM DTT supplemented with 25 mM methionine. Scale bar = 1 mm. Quantification of different developmental stages of N2 (B) and metr-1(ok521) (C) animals after 5 days of hatching at 20°C on E. coli OP50 diet containing 10 mM DTT, 10 mM DTT supplemented with 10 mM methionine, and 10 mM DTT supplemented with 25 mM methionine (n = 3 biological replicates; animals per condition per replicate >60). (D) Representative images of sams-1(ok2946) animals grown on E. coli OP50 diet containing 0 mM DTT, 10 mM DTT, 10 mM DTT supplemented with 25 mM methionine, and 10 mM DTT supplemented with 50 nM vitamin B12. The animals were grown for 5 days on the plates containing methionine and for 3 days under all other conditions. Scale bar = 1 mm. (E) Fraction L4 or adult sams-1(ok2946) animals grown on E. coli OP50 diet containing 0, 10, and 10 mM DTT supplemented with 25 mM methionine, and 10 mM DTT supplemented with 50 nM vitamin B12. The animals were grown for 5 days on the plates containing methionine and for 3 days under all other conditions. ***p < 0.001 via the t-test (n = 3 biological replicates; animals per condition per replicate >50). (F) Relative S-adenosylmethionine (SAM) levels in N2 and rips-1(jsn11) animals grown on E. coli OP50 diet containing 0 mM DTT until the young adult stage, followed by incubation on E. coli OP50 diet containing 0 or 10 mM DTT for 12 hr. **p < 0.01 via the t-test. n.s., nonsignificant (n = 3 biological replicates). (G) Representative images of N2, metr-1(ok521), and sams-1(ok2946) animals after 4 days of hatching at 20°C on E. coli OP50 diet containing 10 and 10 mM DTT supplemented with 80 mM choline. Scale bar = 1 mm. (H) Quantification of different developmental stages of N2, metr-1(ok521), and sams-1(ok2946) animals after 4 days of hatching at 20°C on E. coli OP50 diet containing 10 and 10 mM DTT supplemented with 80 mM choline (n = 3 biological replicates; animals per condition per replicate >50).

Figure 5—source data 1. Methionine and choline supplementation alleviate dithiothreitol (DTT) toxicity.