(A) Representative images of wild-type N2 C. elegans after 72 hr of hatching at 20°C on E. coli OP50 diet containing 0, 10, and 10 mM DTT supplemented with 50 nM vitamin B12. Scale bar = 1 mm. (B) Fraction L4 or adult wild-type N2 C. elegans after 72 hr of hatching at 20°C on E. coli OP50 diet containing 0, 10, and 10 mM DTT supplemented with 50 nM vitamin B12. p values are relative to 0 mM DTT. ***p < 0.001 via the t-test. n.s., nonsignificant (n = 3 biological replicates; animals per condition per replicate >80). (C) Representative images of metr-1(ok521) and mmcm-1(ok1637) animals after 72 hr of hatching at 20°C on E. coli OP50 diet containing 0 mM DTT, 10 mM DTT, and 10 mM DTT supplemented with 50 nM vitamin B12. Scale bar = 1 mm. (D) Fraction L4 or adult metr-1(ok521) and mmcm-1(ok1637) animals after 72 hr of hatching at 20°C on E. coli OP50 diet containing 0, 10, and 10 mM DTT supplemented with 50 nM vitamin B12 (n = 3 biological replicates; animals per condition per replicate >60). (E) Representative images of metr-1(ok521) and mmcm-1(ok1637) animals after 72 hr of hatching at 20°C on C. aquatica DA1877 and P. aeruginosa PA14 gacA mutant diets containing either 0 or 10 mM DTT. Scale bar = 1 mm. (F) Fraction L4 or adult metr-1(ok521) and mmcm-1(ok1637) animals after 72 hr of hatching at 20°C on C. aquatica DA1877 and P. aeruginosa PA14 gacA mutant diets containing either 0 or 10 mM DTT. p values are relative to the corresponding 0 mM DTT condition for each mutant. ***p < 0.001 via the t-test. n.s., nonsignificant (n = 3 biological replicates; animals per condition per replicate>60).
Figure 2—source data 1. Vitamin B12 alleviates dithiothreitol (DTT) toxicity via methionine synthase.