Figure S3. Developmental and spatial expression patterns of
cec-1,
cec-6 and
cec-3.(A) Relative expression levels (dcpm, depth of coverage per base per million reads) from published RNA-Seq data (BOECK et al.2016). Embryogenesis time course represents a unified average of 2-3 timepoint replicates. L1-L4, larval stages 1-4; YA, youngadult; L4 Soma, L4 stage from
glp-1(
q224) mutant animals; Gonad (ad), germlines dissected from adult animals. (B) Relativeexpression levels (transcripts per million) in several cell types from published single-cell combinatorial indexing RNA sequecing(sci-RNA-seq) of L2 animals (CAO et al. 2017).
cec-6 and
cec-3 expression is enriched in the germline, whereas
cec-1 expressionis enriched in somatic tissues. Representative germline-enriched (
mes-4) and neuronal (
rgef-1) transcripts are shown as specificitycontrols. (C) Scheme for generating knock-in epitope-tagged strains using CRISPR/Cas9 gene editing with (right) or without (left)a floxed selection cassette for pharyngeal GFP (Pmyo2::GFP) and G418 resistance (NeoR). (D) Expression of CEC-6::GFP::3xHAin the primordial germ cells (Z2 and Z3, arrowheads) of L1 animals. Z2 and Z3 are identified by immunofluorescence using antiPGL-1 (P granule abnormality protein 1, DSHB mAb K76 (STROME and WOOD 1983) and CEC-6 is detected by anti-HAimmunofluorescence. Scale bar, 20um.