Figure 5(A) qRT-PCR of acs-2, ech-1.1 and fat-5 mRNA expression in 5-day-old mutant nematodes (Mann Whitney-U test, n=6 biological replicates). Data information: bars represent mean±SEM Across experiments, p value summary is *p<0.05, **p < 0.01, ***p<0.001, ****p<0.0001. (B) Simplified schematic of NHR-49/NHR-80 participation to fatty acid beta-oxidation. Green dotted arrows indicate potential transcriptional regulation by NHR-49/NHR-80. In red, enzymes that have been tested in our epistatic analysis. MUFAs is monounsaturated fatty acids, VLCFAs indicates very long chain fatty acid and MCFAs is medium-chain fatty acid. (C) qRT-PCR of nhr-49 expression in wt nematodes grown on control and nhr-49 RNAi expressing bacteria (unpaired t-test, n=3 biological replicates). Data information: bars represent mean±SEM Across experiments, p value summary is *p<0.05, **p < 0.01, ***p<0.001, ****p<0.0001. (D) Lifespan assay of wt nematodes exposed to control, nhr-80 and nhr-49 RNAi. Data information: , the median lifespan ± SEM across replicates is reported underneath the graphs, and additional information (e.g., n numbers) (E-F) Representative survival curves of gas-1(fc21) and gas-1(fc21); cest-2.2 O/E nematodes grown (E) control vs nhr-49 RNAi and (F) control vs nhr-80 RNAi. Data information: the median lifespan ± SEM across replicates is reported underneath the graphs, and additional information (e.g., n numbers) Certain lifespans were run at the same time, but representative figures were split for the sake of clarity (G) qRT-PCR of fat-5 mRNA expression in wt nematodes grown on control and fat-5 RNAi expressing bacteria (unpaired t-test, n=3 biological replicates). Data information: bars represent mean±SEM; Across experiments, p value summary is *p<0.05, **p < 0.01, ***p<0.001, ****p<0.0001. (H-I) Lifespan assay of (H) wt and (I) gas-1(fc21) vs gas-1(fc21); cest-2.2 O/E nematodes grown on control and fat-5 RNAi bacteria. Data information: , the median lifespan ± SEM across replicates is reported underneath the graphs, and additional information (e.g., n numbers) Certain lifespans were run at the same time, but representative figures were split for the sake of clarity (J-K) Representative survival curves of (J) wt and (K) gas-1(fc21) vs gas-1(fc21); cest-2.2 O/E nematodes fed with control and RNAi against mdh-2 (for wt) and acox-1.1. Data information: the median lifespan ± SEM across replicates is reported underneath the graphs, and additional information (e.g., n numbers) Certain lifespans were run at the same time, but representative figures were split for the sake of clarity (L-M) qRT-PCR of (L) ech-1.1 and acox-1.1 deficient (M) acs-2 mRNA expression in wt nematodes grown on control and RNAi expressing bacteria (unpaired t-test, n=3 biological replicates). Data information: bars represent mean±SEM Across experiments, p value summary is *p<0.05, **p < 0.01, ***p<0.001, ****p<0.0001. (N-P) Lifespan assays of (N) wt and (O-P) gas-1(fc21) vs gas-1(fc21); cest-2.2 O/E mutant nematodes grown on control, ech-1.1 and acs-2 RNAi expressing bacteria. Data information: the median lifespan ± SEM across replicates is reported underneath the graphs, and additional information (e.g., n numbers) Certain lifespans were run at the same time, but representative figures were split for the sake of clarity