Figure 1. Genomic location, structure, and expression of
rncs-1. (A) The X chromosome region that encodes the 847-nt
rncs-1 gene is shown. Gray boxes, coding exons; white boxes and arrows, noncoding exons; connecting lines, introns. The 815-bp deletion in
rncs-1(
tm1632) is indicated. Regions used as promoter in Prncs-1::GFP and as rescue fragment in
rncs-1 rescue and overex- pressing lines are marked. (B) Secondary structure of mature
rncs-1 RNA as predicted by mfold and biochemical methods (nuclease probing; data not shown). Asterisks, GU pairs; dots, mismatches and bulges; gray arrowhead, exon- exon junction; numbering, nucleotide relative to transcription start. (C) Northern blot of
rncs-1 in total RNA from wild-type embryos, larval stages (L1-L4), young adults, arrested L1s, and dauer larvae purified from exhausted liquid culture. Blot was rehybridized with probe for 18S ribosomal RNA (rRNA) as a loading control. Data are representative of multiple blots (n = 2-4, depending on stage). (D-F) GFP expression from putative
rncs-1 upstream regulatory and promoter sequences. (D) Comma stage embryo with fluores- cence in the developing midgut (dmid) and cells of developing hypodermis (dh). (E and F) Adult with Prncs-1::GFP expression in the midgut (mid), cells of the head hypodermis (hh), tail hypodermis (th), Hyp 7 syncytium (Hyp 7), and vulval epithelium (ve). GFP expression is absent in seam cells (sc). Focal planes are shown at Bottom Right. Because transgenes are often silenced in the germ line, the lack of GFP expression in the germ line is not necessarily indicative of lack of expression. Multiple transgenic lines showed the same expression pattern. (G) Northern blot of
rncs-1 in total RNA of wild-type hermaphrodites (H) and males (M); 18S rRNA, loading control. (H) Quantification of relative RNA levels in four independent samples of males and hermaphrodites.
rncs-1 levels were determined by Northern blotting;
let-413 and
eps-8 levels were quantified by qRT-PCR. Error bars, SEM; *, P < 0.05, t test.