Extended Data Fig. 2: MIC modulates activity of a nuclear hormone receptor DAF-12/FXR.
a, mRNA of lysosomal and mitophagy genes, day-1 WT(N2) and daf-12(rh61rh411) (20 °C). Assay was independently repeated four times. Graph show mean ± s.d. (p-values via two-tailed unpaired t-test). b, WT(N2) and daf-12(rh61rh411), day-1 (20 °C); Left: lysotracker intensity, (n = 311 and n = 325 lysosomes), p < 0.0001. Right: lysosomal area, (n = 311 and n = 321 lysosomes), p = 0.0013. Assay was independently repeated three times with ≥15 animals/repeat. (P-value via two-tailed unpaired t-test). Scale: 10 µm. c, WT(N2) and daf-12(rh61rh411) 24 h post-DMSO/MIC treatment, day-2 (20 °C); lysotracker intensity, Left: (n = 272 and n = 617 lysosomes), p < 0.0001; Right: (n = 325 and n = 477 lysosomes), p < 0.0001. Assay was independently repeated three times with ≥15 animals/repeat. (p-value via two-tailed unpaired t-test). d. mRNA of lysosomal genes, day-2 WT(N2) and hlh-30(tm1978) (20 °C). Assay was independently repeated five times. Graph show mean ± s.e.m. (p-values via two-tailed unpaired t-test). Right: lysotracker intensity, hlh-30(tm1978) 24 h post-DMSO/MIC treatment, day-2 (20 °C), (n = 447 and n = 156 lysosomes), p < 0.0001. Assay was independently repeated three times with ≥15 animals/repeat. (P-value via two-tailed unpaired t-test). The box plots (b, c and d) represent the 25th–75th percentiles, the line depicts the median and the whiskers show the min–max values. e, DAF-12 activity post-dafachronic acid (ligand) doses quantified by one-hybrid reporter assay. Graph show mean ± s.d. Assay was independently repeated four times, except for ligand 50 nM that is, two, Ligand200 (p = 0.0045) and Ligand500 (p = 0.0003), (P-value via one-way ANOVA with Tukey’s multiple comparisons test). f, Schematic depicts dafadine A (DFA) action. DFA induces HLH-30 expression. HLH-30::GFP (MAH235), day-2 animals post-DFA/DMSO treatment (20 °C) probed with GFP antibody, Actin (ACT) as loading control. Assay was independently repeated two times. g, mRNA of lysosomal genes, day-2 WT(N2) and daf-12(rh61rh411) (20 °C). Assay was independently repeated two times. (p-values via two-tailed unpaired t-test).
