0001). skn-1(zu169) −/− fed GD1 showed a 69% increase

in mean life span compared to mutants fed OP50 (b, p < .0001). Data were subjected to one-way ANOVA with Fisher’s test at a significance level of p < 0.05. A growing body of evidence indicates that the increased life span of C. elegans fed the GD1 diet is not due to the lack of Q per se. C. elegans clk 1 mutants also show enhanced life span in response to the GD1 diet [17]. The clk 1 mutants lack Q but continue to produce rhodoquinone, an amino-isoprenylated quinone involved in GSK3235025 purchase anaerobic respiratory metabolism, as well as demethoxy-Q, the penultimate intermediate in Q biosynthesis [23, 24]. To determine whether the GD1 diet would also act to extend life span of a C. elegans mutant with an earlier defect in the Q biosynthetic pathway, we tested the effects of this diet on two C. elegans coq 3 mutants. COQ-3 is an O-methyltransferase required for mTOR inhibitor review two steps of Q biosynthesis: the first O-methylation step precedes formation of the quinone ring, and the second O-methylation step is the final step, producing Q [25]. C. elegans coq 3 mutants have more severe phenotypes than the clk 1 mutants [20, 26]. The coq 3 mutant worms respond to the GD1 E. coli diet when maintained on the diet either from time

of hatching (Figure 2A), or when the diet is provided to the mutants upon reaching the L4 larval stage (Figure 2B). These results indicate that the GD1 diet imparts life span extension even to worm mutants with severe early defects in Q biosynthesis, and hence its effects are independent Carbohydrate of worm Q content. Figure 2 Q deficient worms respond to GD1 diet. (A) Wild-type (squares), coq-3(ok506) −/− (circles) and coq-3(qm188) −/− (diamonds) were fed either OP50 (black) (N2, n = 529; coq-3(ok506) −/−, n = 119; coq-3(qm188) −/−, n = 259) or GD1 (grey) (N2, n = 225; coq-3(ok506) −/−, n = 102; coq-3(qm188) −/−, n = 141) from the hatchling

stage and assessed for survival. Asterisks designate: A significant PLX3397 order increase in mean life span of N2 fed GD1 compared to OP50: 37% (p < .0001); Increase in mean life span of coq-3(ok506) −/− fed GD1 compared to N2 fed OP50: 58% (p < .0001); and Increase in mean life span of coq-3(qm188) −/− fed GD1 compared to N2 fed OP50: 74% (p < .0001). (B) Wild-type (squares) and coq-3(ok506) −/− (circles) were fed OP50 (black) until the L4 larval stage and then subsequently fed either OP50 (black) (N2, n = 63; coq-3(ok506) −/−, n = 84) or GD1 (grey) (N2, n = 55; coq-3(ok506) −/−, n = 53) and assessed for survival. Increase in mean life span of N2 worms fed GD1 compared to N2 fed OP50: 75% (p < .0001). Increase in mean life span of coq-3(ok506) −/− fed GD1 compared to N2 fed OP50: 113% (p < .0001). Data were subjected to one-way ANOVA with Fisher’s test at a significance level of p < 0.05.