Extended Data Fig. 3: MIC-induced mitophagy enhances mitochondrial health.
a. Image depicts co-localization (arrowheads) in day-2 reporter animals treated with DMSO, MIC, or FCCP at 20 °C, expressing mCherry::LGG-1 and mitochondrial GFP. FCCP is used as positive control. Right: Spearman’s co-localization coefficient. Assay was independently repeated four times. Graph show mean ± s.d. MIC (p = 0.0464) and FCCP (p = 0.0003), (p-value via two-tailed unpaired t-test). Scale: 10 µm. b. mRNA of mitophagy genes in day-2 WT(N2) and daf-12(rh61rh411) mutants 24 h post DMSO or MIC (20 °C). Assay was independently repeated four times. Graph show mean ± s.d. daf-12 vs WT, for dct-1 (p = 0.0059) and drp-1, (p = 0.0476), WT (DMSO vs MIC), for dct-1 (p = 0.0071) and for drp-1 (p = 0.0326), (p-value by unpaired t-test). c. mRNA of mitophagy genes in day-2 WT(N2) and hlh-30(tm1978) mutants 24 h post DMSO or MIC (20 °C). hlh-30 vs WT, for dct-1 (p = 0.0099), pink-1 (p = 0.0277). WT (DMSO vs MIC), for dct-1 (p = 0.0004) and hlh-30 (DMSO vs MIC) for pink-1 (p = 0.0163). Assay was independently repeated three times. Graph show mean ± s.d., (p-value by unpaired t-test). d. Quantification of mitophagy in muscle-specific mito-rosella reporter upon RNAi treatment, performed at day-2, 24 h post DMSO/MIC (20 °C). Assay was independently repeated three times (n = 45, 44, 34 and 26 animals). Graph show mean ± s.d. control RNAi, MIC vs DMSO (p = 0.0020), (p-value via one-way ANOVA with Tukey’s multiple comparison test). e. Survival of DMSO/MIC treated day-5 pdr-1(gk448) mutants 5 h (20 °C) post-rotenone. Assay was independently repeated three times. Graph show mean ± s.d., p = 0.0226 (p-value via two-tailed unpaired t-test). f. Oxygen consumption rate, basal (B) and max (M) of day-2 WT(N2) and hlh-30(tm1978) mutants 24 h post DMSO/MIC (20 °C). Assay was independently repeated four times. Graph show mean ± s.e.m. (B vs M) for; WT-DMSO (p = 0.0169), WT-MIC (p < 0.0001), hlh-30-DMSO (p = 0.0269), hlh-30-MIC (p = 0.0005); (max-WT-DMSO vs max-WT-MIC, p < 0.0001), (max-WT-MIC vs max-hlh-30-MIC, p < 0.0001), (p-value via two-way ANOVA with Sidak’s multiple comparison test).
