Generation of transgenic strains and strain list for C. elegans

To generate transgenic lines for localization, rescue experiments, and expression patterns, we generated the following transgenic animals via microinjections.

OIK1042 turEx21[arl-13p::wdr-31 (T05A8.5)::GFP::unc-54 3′UTR +rol-6}; T05A8.5(syb1568)II., elmd-1(syb630) II, rpi-2(K08D12.2)(ok1863) IV. (1 ng).

OIK1044 N2;turEx23[elmd-1p::GFP::elmd-1 (C56G7.3)::unc-54 3′UTR +rol-6} (5 ng).

OIK1045 N2;turEx24[arl-13p::GFP::elmd-1 (C56G7.3)::unc-54 3′UTR +rol-6} (5 ng).

OIK1046 N2;turEx25[elmd-1p(C56G7.3)::GFP::unc-54 3′UTR +rol-6} (50 ng).

OIK1047 N2;turEx26[wdr-31 (T05A8.5)p::GFP::unc-54 3′UTR +rol-6} (50 ng).

The rol-6 plasmid (50 ng/μl plasmid pRF4) was co-injected as the co-transformation marker. In brief, the plasmids were delivered by microinjections into the gonads of 1-d adult worms. Worms were initially transferred onto a 2.5% agarose pad before (Halocarbon oil, 9002-83-9; Sigma-Aldrich), followed by microinjection. The microinjection was done using a Zeiss Axio Vert.A1 inverted microscope with DIC optics coupled with a Narishige Micromanipulator MMO-4. We next manually inspected the plates to find successful transgenic animals.

N2; FX30333; wdr-31(T05A8.5)(tm10423)II.; OIK393 T05A8.5(tur003)II.; PHX1568 T05A8.5(syb1568)II.; nphp-4(tm925)V.; mks-6(gk674)I.; PHX630, elmd-1(syb630) III.; RB1550 rpi-2(K08D12.2)(ok1863) IV.; MX52, bbs-8(n x 77) V. We obtained wdr-31(T05A8.5)(tm10423)II. (FX30333) mutant allele, which has a 160-bp deletion, causing a frameshift, from the National Bioresource Project. The wdr-31(T05A8.5)(tm10423)II. was outcrossed to WT four times. The Caenorhabditis Genetics Center (CGC) provided the RB1550 rpi-2(K08D12.2)(ok1863) mutant, and the rpi-2(ok1863) allele contains 1,143-bp deletion that removes a large segment of exon III, exon IV, and some portions of exon V. T05A8.5(syb1568)II. has 1,888 bp deletions, deleting all exons except exon I (Fig S3B). Sunybiotech created an independent null allele of elmd-1 via CRISPR–Cas9. The PHX630, elmd-1(syb630) III. mutant contains 1,784 bp deletions, where except for exon I, all exons were removed (Fig S2B). elmd-1(syb630) III. were outcrossed to WT two times.

GOU2162 che-3(cas443[gfp::che-3]) I; xbx-1(cas502[xbx-1::tagRFP]) V; GOU2362 ift-74(cas499[ift-74::gfp]) II.; EJP76 vuaSi15 [pBP36; Posm-6::osm-6::eGFP; cb-unc119(+)] I; unc-119(ed3) III; osm-6(p811) V; N2;lqIs2[osm-6::gfp], N2;ejEx[osm-3::GFP + pRF4]; N2;ejEx[kap-1::gfp+pRF4]; EJP81.

vuaSi24 [pBP43; Pift-140::ift-140::mCherry; cb-unc-119(+)]II; unc-119(ed3) III; ift-140(tm3433) V; jhuEx [ift-140::GFP+pRF4]; Ex[rpi-2::GFP+ xbx-1::tdTomato+pRF4]; MX76 dpy-5(e907); nxEx(bbs-7::gfp+dpy-5 (+)).

PHX1180, wdr-31(syb1180 [wdr-31(T05A8.5)::GFP]); PHX4934 rpi-2(syb4934) [rpi-2::mCherry]; OIK1042 N2;turEx21[arl-13p::wdr-31 (T05A8.5)::GFP::unc-54 3′UTR +rol-6} (5 ng); OIK1044 N2;turEx23[elmd-1p::GFP::elmd-1 (C56G7.3)::unc-54 3′UTR +rol-6} (5 ng); OIK1045 N2;turEx24[arl-13p::GFP::elmd-1 (C56G7.3)::unc-54 3′UTR +rol-6} (5 ng); OIK1046 N2;turEx25[elmd-1p(C56G7.3)::GFP::unc-54 3′UTR +rol-6} (50 ng); OIK1047 N2;turEx26[wdr-31 (T05A8.5)p::GFP::unc-54 3′UTR +rol-6} (50 ng); N2;Ex[mks-2::GFP + tram-1::tdTOMATO + pRF4]; MX1409 N2; nxEx785[tax-4:gfp+ Posm-5::xbx-1::tdTomato + rol-6(su1006)]; vuaSi21[pBP39; Pmks-6::mks-6::mCherry; cb-unc-119(+)]II; MX2418 N2;nxEx1259[pbbs-8::PLC-delta PH::GFP;MKSR-2::tdTomato; coel::GFP]; PY8847 oyIs65[str-1p::mcherry]; Ex[str-1p::nphp-4::gfp, unc122p::dsRed]. All PHX strains were generated using CRISPR/Cas9 by Sunybiotech. The list of extensive transgenic and mutant strains is provided in Table S4.

Table S4. Both the strain names and their genotypes are displayed. The lab sources or those who created the strain are displayed.