Figure S1. Related to Figures 1, 2, 3, 5, and 6. MORC-1 is expressed in somaticnuclei, characterization of E39A variant,
morc-1 does not regulate siRNAaccumulation, RNA-seq and ChIP-seq strategy overview. (A) MORC-1::3xFlag isexpressed in intestinal nuclei shown by anti-Flag immunofluorescence and DAPI stainingof worms expressing
morc-1::3xflag at the endogenous
morc-1 locus and grown for twogenerations on empty vector RNAi or on
morc-1 RNAi. (B) Western blot of wormsexpressing
morc-1(WT)::3xflag and
morc-1(E39A)::3xflag grown on vector or morc-1RNAi using anti-Flag antibody. Anti-H3 antibody was used as a loading control. Asteriskdesignates a higher molecular weight band that is eliminated by
morc-1 RNAi. This mayindicate a post-translational modification or alternative isoform of MORC-1. Quantificationof both bands by LI-COR Odyssey Fc is shown in (C). (D) Fertility of designated genotypeat F6 and F10 generations at 25C. Mean fertility ± SD of 12 worms per genotype pergeneration is shown.
morc-1(-) mutants are sterile at F6,
morc-1(E39A)::3xflag mutantsare essentially sterile at F10, and
morc-1(WT)::3xflag worms have wildtype fertility. (E)
morc-1(E39A)::3xflag mutants are resistant to
lir-1 RNAi.
morc-1(WT)::3xflag wormshave wildtype sensitivity to
lir-1 RNAi. Bars indicate percentage of worms arrested ordead as a mean of three biological replicates ± SD;
n100 (F)
morc-1(E39A)::3xflag isdefective for RNAi inheritance. On gfp RNAi, silencing of gfp is completely penetrant, butthere is a mild defect in gfp silencing in the F1 generation. In
morc-1(-) mutants, themajority of F1 worms express GFP. Silencing is completely penetrant in P0 and F1generations in
morc-1(WT)::3xflag and wildtype worms. Bars indicate percentage ofworms expressing GFP as mean of three biological replicates ± SD;
n50. (G) Levels ofanti-gfp siRNAs in adult worms grown on gfp RNAi and in F1 and F2 generations grownon OP50. siRNA levels measured by TaqMan qRT-PCR, normalized to U18, and shownin arbitrary units as mean of two technical replicates ± SD. (H) Sample collectionschematic for RNA-seq and ChIP-seq studies.