Supplementary Figure 4: DAMT-1 promotes global induction of 6mA upon mitochondrial stress.

a, Genotyping of damt-1 mutant. b, Representative fluorescent images of hsp-6p::gfp or damt-1; hsp-6p::gfp worms fed on control or spg-7 RNAi. Scale bar: 100 μm. c, Genotyping of damt-1 germline rescued worms. d, Intergenerational inheritance tested in damt-1; pie-1p::damt-1 animals. e, Quantification of global 6 mA level in (Fig. 3c) by UHPLC–QqQ-MS/MS. f, Dot blot of untreated wild-type worms, or worms treated with 1.92 μM antimycin, 90 μM TTFA, 4.62 μM NaN₃, or 0.64 μM tunicamycin. g, Dot blot of sequential tunicamycin-treated wild-type animals. h, Total RNA and genomic DNA were extracted and tested with DNA 6 mA antibody on dot blot. The result shows that the signals on dot blot probed with DNA 6mA antibody is not due to any contamination of RNA m6A. i, Quantification of total RNA m6A level upon antimycin treatment by UHPLC–QqQ-MS/MS. j, DNA dot blot of wild-type animals fed on dam- dcm- E.coli and treated with or without antimycin. k, Schematic representation of CRISPR–Cas9-mediated knockout of damt-1 in worms. l, Dot blot (top) and intergenerational inheritance (bottom) of damt-1 Cas9 knockout animals. m, in vitro methyltransferase assay tested in nuclear lysates from untreated or antimycin-treated wild-type animals or damt-1 mutants. Worms for dot blot were collected when they reached L4 stage. Data in a-c, f-h, j and l represent results from three independent assays. n = 3 biologically independent experiments (d,i,l,m), >100 animals per experiment (d, l bottom), ~ 100,000 animals per experiment (i,m). Graph data are presented as mean ± s.e.m. Statistical analysis was performed by paired (d,l) or unpaired (i,m) two-tailed t-test; ns, not significant, P > 0.05. Statistical source data are provided in Supplementary Table 7. Uncropped dot blot figures are presented in Supplementary Figure 6.