Animals were maintained under standard procedure with nematode growth media (NGM) plates, unless otherwise stated. Bristol strain N2 was used as WT, and Hawaiian strain CB4856 was used for the linkage analysis of the mutants (59, 60). hir-1 null alleles were generated by CRISPR to induce double-stranded breaks, and subsequent nonhomologous end joining caused a deletion of hir-1. Feeding RNAi was performed as previously described (61). Transgenic strains were generated by germline transformation as described (62). Transgenic constructs were coinjected with dominant unc-54p::mCherry or myo-2::mCherry markers, and stable extrachromosomal lines of mCherry+ animals were established. Transgenic strains used were dmaIs1[comt-5p::GFP], dmaIs1; egl-9(sa307), dmaIs1; hif-1(ia4), dmaIs1; hir-1(dma51), dmaIs1; hir-1(tm4098), dmaIs1; hir-1(tm3911), dmaIs1; hir-1(dma101), dmaIs1; hir-1(dma101); dmaEx113[dpy-7p::hir-1(+); myo-2::mCherry], dmaIs1; hir-1(dma101); dmaEx113[ric-19p::hir-1(+); myo-2::mCherry], dmaIs1; dpy-3(dma22), dmaIs1; dpy-2(dma52), dmaIs1; perl-1(dma53), dmaIs1; perl-1(dma236), dmaIs1; dma11, dmaIs1; dma11; mdt-15(dma54), dmaIs1; dma11; nhr-49(dma55), dmaIs1; nhr-49(nr2041), dmaIs1; hir-1(dma101); nhr-49(nr2041), dmaIs1; hir-1(dma101); hif-1(ia4), dmaIs1; dpy-3(ok2263), dmaIs1; dpy-3(ok2263); nhr-49(nr2041), dmaIs1; hir-1(dma101); dpy-3(ok2263), dmaIs1; hir-1(dma101); dpy-3(ok2263); nhr-49(nr2041), dmaIs1; nhr-49(et7), dmaIs1; mdt-15(et14), dmaEx114[rpl-28p::nhr-49::Venus; myo-2::mCherry], dmaEx79[hir-1p::GFP; unc-54p::mCherry], dmaEx121[rpl-28p::GFP::hir-1; myo-2p::mCherry], dmaEx229[hir-1p::hir-1::Flag; unc-54p::mCherry], dmaEx229; dpy-3(ok2263), dmaEx206[snr-2p::let-756::HA; unc-54p::mCherry], dmaEx211[dpy-3p::dpy-3::FLAG; unc-54p::mCherry], dmaEx257[snr-2p::let-756::HA], DMS1101_dmaIs1; dpy-18(e364), dmaEx258[dpy-7p::dpy-3::Venus; unc-54p::mCherry], dmaEx258; hir-1(dma101), kaIs12[col-19::GFP], kaIs12; dpy-18(e364), kaIs12; hir-1(dma101), and kaIs12; hir-1(dma101); nhr-49(nr2041).

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