Fig. 2

Analysis of combinatorial treatments on C. elegans gas-1(fc21) complex I mutant worms’ lifespan. Lifespan analyses were performed at 20°C in gas-1(fc21) worms treated from the L1 early larval development stage with one of 11 randomly selected triple therapy combinations of three treatment classes that had previously been found to each significantly increase to variable degrees these complex I mutant worms’ lifespan (Supplementary Material, Figs. S2 and S3). All drug-treated lifespan analyses were performed relative to buffer-only treated gas-1(fc21) and N2 (WT) worms. (A-B) Lifespan results are shown for the 2 of 11 tested combinatorial therapy regimens that synergistically rescued gas-1(fc21) lifespan, even beyond that of WT worms. Key indicates number of worms studied across two biological replicate experiments per condition (n). Statistical analysis was performed by log-rank (Mantel-Cox) test comparing N2 (buffer only) or gas-1(fc21) worms treated with the indicated combination relative to buffer-only treated gas-1(fc21) worms. Subsequent analyses indicated that apparent efficacy of combination #6 (resveratrol + folinic acid + cysteamine bitartrate) was driven by resveratrol effect, without further synergy from additive therapies tested. (C) Glu + NA + NAC in triple or pairwise combinations showed synergistic rescue effect of gas-1(fc21) worms’ median lifespan relative to each individual component(s). Results convey median lifespan normalized to buffer-only exposed gas-1(fc21) worms. Detailed results are shown in Supplementary Material, Fig. S4.