0001 a, b, c, d, e, f, identify cohorts from the same experiment. Within each cohort data were check details subjected to One-Way ANOVA analyses with Fisher’s test at a significance of 0.05. (p-values are compared to the condition in bold text for a given cohort). Worms fed GD1 are more thermotolerant and resistant to juglone treatment BMN 673 supplier mutants of C. elegans with life span extension often show enhanced resistance to thermal and oxidative stress
, suggesting that worms fed the GD1 diet would also demonstrate stress resistance. Juglone is a quinone that imposes both oxidative and electrophilic stress [27, 28]. Juglone penetrates the worm cuticle and has been used to select for oxidative stress-resistant mutants . As shown in Figure 4A, worms fed GD1 from the hatchling stage display improved survival following exposure to 250 μM juglone, as compared to similarly treated worms fed OP50. It is unlikely that the improved worm survival is due to hypersensitivity C646 purchase of GD1 E. coli to juglone treatment because the GD1 E. coli were actually more resistant to juglone treatment than OP50 E. coli (Additional file 1). Similarly, worms fed GD1 are more thermotolerant at the L4 stage
compared to worms fed OP50 (Figure 4B). Figure 4 GD1-fed worms are more resistant to juglone treatment and show enhanced thermotolerance. (A) Wild-type N2 worms were fed OP50 or GD1 from the hatchling stage. L4 larval worms were placed in a drop of S-media containing either Rutecarpine 250 μM juglone or an equal amount of ethanol vehicle control for 20 min. Worms were washed onto OP50 plates to recover and assayed for survival 18 h later. Black bar: OP50, grey bar: GD1; Asterisk indicates p-value = 0.0003 determined with Student’s t-test when compared to the OP50 + juglone condition. (B) Wild-type N2 worms were fed OP50 or GD1 from the hatchling stage. L4 larval worms were incubated at 35°C and survival was assessed at each indicated time point. Black line: OP50, grey line: GD1. Asterisks indicate p-values determined with Student’s t-test for comparisons between GD1 and OP50 at the designated time
points: (7 h) 0.003; (9 h) 0.0013; (10 h) 0.0001; (11 h) 0.017. Excreted components present in GD1 E. coli spent media are not responsible for life span extension Previous studies have shown that E. coli mutants with defects in the ubiA gene, required for Q biosynthesis, excrete large amounts of D-lactic acid in the spent media . We found that the spent media of both GD1 and GD1:pBSK E. coli contain millimolar quantities of D-lactic acid (Figure 5A). In contrast, the spent media collected from cultures of OP50 contain only 10–20 μM D-lactic acid, similar to the concentration observed in LB media alone. Similarly, rescued GD1 cells containing a wild-type copy of ubiG produce very low levels of D-lactic acid, indicating that excretion of D-lactic acid by the GD1 E. coli is due to the loss of Q biosynthesis.