Figure 2
Validation of FA and AA analysis in C. elegans with deficient lipid or AA metabolism. (a) Schematic representation of polyunsaturated fatty acid (PUFA) synthesis pathway in C. elegans. mdt-15 is a key regulator in this pathway that controls the activity of stearoyl-CoA desaturases fat-6 and fat-7. (b) N2 worms exposed to mdt-15 RNAi accumulated C18:0 and had reduced levels of C18:1 and PUFAs. Bar graphs are expressed as mean ± SD; Significance was calculated using Student’s t-test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001. (c) FA composition in mdt-15 RNAi bacteria was similar to that in the control RNAi bacteria. No PUFAs were detected in either RNAi bacteria. (d) Schematic representation of branched-chain amino acid (BCAA) catabolism. Branched-chain amino acid aminotransferase 1 (bcat-1) catalyzes the first reaction of this pathway. (e) Accumulation of BCAAs valine, leucine and isoleucine was detected in rrf-3(pk1426) worms treated with bcat-1 RNAi. Worms grown on plates supplemented with 20 mM BCAAs accumulated BCAAs, which was more pronounced in combination with bcat-1 RNAi. Bar graphs are expressed as mean ± SD, Significance was calculated using One-way ANOVA; *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001. (f) BCAAs levels were similar in control and bcat-1 RNAi bacteria, but accumulated in BCAA-supplemented bacteria. See also Supplementary Fig. S2.
