Extended Data Fig. 3: Characterization of mitochondrial defects and metabolites in C. elegans mutants affecting propionate breakdown.
From: A feedback loop engaging propionate catabolism intermediates controls mitochondrial morphology
a, Representative images of hypodermal mitochondria in hphd-1(ok3580) and dhgd-1(tm6671) mutants grown on E. coli OP50 without or with addition of VB1, VB6, and VC. Bars, 5 μm. b-d, Left: Quantification of Val (b), Ile (c) and Leu (d) in N2, dhgd-1(tm6671) and hphd-1(ok3580) animals fed on E. coli OP50. Right: Representative chromatograms of each amino acid in the indicated worms. Worms grown on 20 NGM plates (10 cm in diameter) were analyzed for each genotype. n = 4 independent experiments for N2 and dhgd-1(tm6671) and n = 3 independent experiments for hphd-1(ok3580). e, Left: Quantification of KMP (top) and KMV (bottom) in N2, dhgd-1(tm6671) and hphd-1(ok3580) animals fed on E. coli OP50. Right: Representative chromatograms of KMP and KMV in the indicated worms. Worms grown on 20 NGM plates (10 cm in diameter) were analyzed for each genotype. n = 4 independent experiments for N2 and dhgd-1(tm6671) and n = 3 independent experiments for hphd-1(ok3580). f, Representative images of mitochondria in the hypodermis of N2, pcca-1(ok2282) mutants, control RNAi-, mce-1 RNAi- and mmcm-1 RNAi-treated worms fed on E. coli OP50. RNAi was performed using RNAi-competent OP50 E. coli. Bars, 5 μm. g, Representative images of mitochondria in the hypodermis of N2, acdh-1(ok1489), alh-8(ww48), control RNAi-, ech-6 RNAi- and hach-1 RNAi-treated worms fed on E. coli OP50. RNAi was performed using RNAi-competent E. coli OP50. Bars, 5 μm. h, Representative images of mitochondria in the hypodermis of N2, acdh-1(ok1489) and dhgd-1(tm6671) single mutants and acdh-1;dhgd-1 double mutants fed on E. coli OP50. Bars, 5 μm. i. Quantification of propionate in worms with the indicated genotype. Worms grown on 20 NGM plates (10 cm in diameter) were analyzed for each genotype. n = 3 independent experiments. For statistical analyses, P values were determined using one-way ANOVA (i) and two-sided Student’s t-test (b, c, d and e). Individual data points (mean ± s.e.m. and exact P value) are shown. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. Source numerical data are available in source data.
