Picture from Lyssenko NN et al. (2008) BMC Dev Biol "An unexpectedly high degree of specialization and a widespread involvement in ...."

Figure 6. tat-4 reporter expression pattern. The 5' end of the tat-4 locus, location of the deleted region in tm1801 mutants (bar) and structure of the tat-4 expression cassette (A). Reporter expression in the pharyngeal-intestinal valve, intestine and rectal gland cells in 3-fold embryos (B) and in advanced stage larvae (C and D) in the lcIs911.30 line. GFP fluorescence in the uterus (E), during spermatogenesis (F) and in the spermatheca of previously ovulated hermaphrodites (G) in lcIs911.30 animals. Abbreviations: pv - pharyngeal-intestinal valve; r - rectum; rgc - rectal gland cells; sp - spermatheca.

Picture from Lyssenko NN et al. (2008) BMC Dev Biol "An unexpectedly high degree of specialization and a widespread involvement in ...."

Figure 5. tat-3 reporter expression pattern. The 5' end of the tat-3 locus, location of the deleted region in tm1275 mutants (bar) and structure of the tat-3 expression cassette (A). tat-3 reporter expression in the pharyngeal-intestinal valve and in muscle and marginal, but not gland, cells of the pharynx in the lcIs471.12 line (B). GFP fluorescence in the muscle and buccal epithelial cells of the pharynx procorpus and in the XXX cells in the lcIs471.4B line (C). tat-3 reporter signals in the rectum and a tail region (D), seam cells and the hypodermis (E), the DTC (F) and the AC (G) of lcIs471.4B animals. GFP staining of the vulva at the late L4 stage in lcIs881.3.2 larvae (H). Reporter expression in the adult lcIs471.4B vulva (I). Cell labels: AC - anchor cell; DTC - distal tip cell; e1 and 2 - buccal epithelial cells; g1 - gland cell; mc - marginal cell; pm2, 5, 6 and 7 - pharyngeal muscle cells; vul - vulval cells; utse - uterine seam cell; XXX - XXX cells. Abbreviations: hn - hypodermal nucleus; pv - pharyngeal-intestinal valve; sn - seam cell nucleus.

Picture from Lyssenko NN et al. (2008) BMC Dev Biol "An unexpectedly high degree of specialization and a widespread involvement in ...."

Figure 4. tat-2 reporter expression pattern. The 5' end of the tat-2 locus, locations of the deleted regions in tm1634 and tm1773 mutants (bars) and structure of the tat-2 expression cassette (A). Expression of the tat-2 reporter in the lcIs982.4.2 line in embryos (B), in the intestine of L2 larvae (C) and in gland cells of the pharynx, the pharyngeal-intestinal valve, the excretory gland cell and the intestine of a young adult (D). GFP fluorescence in the lcEx982.2.4.5 line in the excretory pore and excretory canal cells (E). Reporter signal outlining the vulF cells in early lcIs982.4.2 L4 larvae (the third frame in the series shows an overlay of the pseudocolored UV image onto the bright light image, both enlarged with re-sampling using Photoshop; dashes extend along the contact surface between the two tissues) (F). tat-2 reporter expression during spermatogenesis (G) and in adult spermatheca (H) of lcIs982.4.2 animals. Abbreviations: eg ' excretory gland cell; epc ' excretory pore cell; g 'pharyngeal gland cell; ga 'gonad arm; ph ' pharynx; pv ' pharyngeal-intestinal valve; oo ' oocyte; r ' rectum; sp ' spermatheca.

Picture from Gracida X et al. (2013) Curr Biol "Fertility and germline stem cell maintenance under different diets requires ...."

(D) Wild-type germ lines show nhr-114 mRNA expression (purple staining) throughout the germ line. nhr-114(gk849) germ lines serve as a negative control and show no nhr-114 mRNA staining. Asterisk indicates the position of the distal end. (E) Section of extruded intestines. From left to right, DIC microscopy image of intact animal focusing on the most anterior part of the intestine. pv: pharyngeal valve (bracket), i: intestine (dotted line). In wild-type nhr-114 mRNA (dark purple) enriches to the nucleoplasm (arrows). nhr- 114(gk849) intestines serve as a negative control and show no nhr-114 mRNA in nuclei. Right images are longer incubation times of different intestines. (F-H) Characterization of intestinal NHR-114 transgenic protein. (F) Immunostaining of extruded intestines of the EV441 strain (n>20 animals). anti-FLAG detects FLAG::NHR-114 in intestinal cells DAPI visualizes nuclei. Anterior (top); posterior (bottom). antianillin (ANI-2) outlines apical membranes. Scale bar 50 um. (G) Nuclear enrichment of FLAG::NHR-114 in intestinal cells. DAPI-positive bacterial remnants outline intestinal lumen. Scale bar 50 um. (H) Anti-FLAG and anti-tubulin immunoblot. FLAG::NHR-114 protein levels remain unchanged upon nhr-114 RNAi knockdown. Lanes 3 and 4 represent two independent experiments. Effectiveness of RNAi knockdown of endogenous nhr-114 was confirmed by the appearance of steriles in non-transgenic siblings.

Picture from Hwang HY et al. (2002) Proc Natl Acad Sci U S A "The Caenorhabditis elegans vulval morphogenesis gene sqv-4 encodes a ...."

Figure 3. SQV-4 protein expression. (A-H) Selected confocal images of SQV-4 Ab staining in multiple planes of focus along the left-to-right axis of the WT animal. Vulval cells with increased SQV-4 expression are indicated. (A-C) SQV-4 expression is increased in cells containing 10 of 22 vulval nuclei in the early L4 stage. The cells strongly stained with SQV-4 Abs dorsal to the vulval cells are in the uterus. The images are arranged from the left-most plane of focus to the right-most plane. (A) Vulval cells containing four nuclei (P5.ppal, P6.paal, P6.pppl, and P7.papl) on the left side of the worm. (B) Two vulval cells (P5.ppp and P7.paa) in the middle plane of the worm. (C) Vulval cells containing four nuclei (P5.ppar, P6.paar, P6.pppr, and P7.papr) on the right side of the worm. (D-H) SQV-4 expression is increased in cells containing 14 of 22 vulval nuclei in the mid-late L4 stage. The images are arranged from the left-most plane of focus to the right-most plane. (D) Vulval cells containing the same four nuclei as in A. (E) Vulval cells containing two of the four dorsal-most nuclei (P6.papl and P6.ppal), which were not stained at an earlier stage. (F) Vulval cells containing the same two nuclei as in B. (G) Vulval cells containing two of the four dorsal-most nuclei (P6.papr and P6.ppar), which were not stained at an earlier stage. (H) Vulval cells containing the same four nuclei as in C. (I) SQV-4 Abs stain oocytes. A row of oocytes in an adult hermaphrodite stained with SQV-4 Abs is indicated with arrows. The oocyte most proximal to the uterus is located at the lower left. (J-K) SQV-4 protein expression in mutants with abnormal vulval development. (J) Confocal image of a lin-12(gf) mutant fixed and stained with SQV-4 Abs. Each pseudovulva (pv) has three nuclei strongly stained with SQV-4 Abs. The functional vulva (v) has six nuclei strongly stained with SQV-4 Abs. Not all staining is shown here. (K) Confocal image of an L4 lin-11(lf) mutant fixed and stained with SQV-4 Abs. Staining by SQV-4 Abs is limited to one cell in this focal plane. Staining is absent in vulval cells in other focal planes (not shown). (Scale bar = 10 um.)

Picture from Gracida X et al. (2013) Curr Biol "Fertility and germline stem cell maintenance under different diets requires ...."

Figure S1. Expression of nhr-114 mRNA and Transgenic Intestinal FLAG::NHR-114 (A-C) Northern blot analysis using antisense nhr-114 probe (described in Figure 1A). (A) In wild-type hermaphrodites, the antisense probe hybridizes to a single product. No signal is present in nhr-114(gk849) mutants, whereas two different products are present in nhr-114(ef24) mutants (green triangles, kb: kilobases). The smaller one is consistent with a 3'end truncation and the larger one is consistent with a 3'end formation defect, such as an alternative choice of polyadenylation sites. 18S rRNA serves as loading control; inf-1 mRNA is an internal control for a germline-enriched gene. (B) nhr-114 mRNA is more abundant in adult hermaphrodites with germ cells than in those without germ cells. Males are him-8(e1489), females are fog-1(253ts). Wild type hermaphrodites (black symbol) with germ lines (+), and without germ lines glp-1(q224ts) hermaphrodites (-). RNA from nhr-114(gk849) hermaphrodites serves as a negative control for the nhr-114 probe. (C) Constant expression of nhr-114 mRNA through development. E=embryo; L1-L4= larval stages; YA=young adult. (D and E) In situ hybridizations using antisense nhr-114 probe. (D) Wild-type germ lines show nhr-114 mRNA expression (purple staining) throughout the germ line. nhr-114(gk849) germ lines serve as a negative control and show no nhr-114 mRNA staining. Asterisk indicates the position of the distal end. (E) Section of extruded intestines. From left to right, DIC microscopy image of intact animal focusing on the most anterior part of the intestine. pv: pharyngeal valve (bracket), i: intestine (dotted line). In wild-type nhr-114 mRNA (dark purple) enriches to the nucleoplasm (arrows). nhr- 114(gk849) intestines serve as a negative control and show no nhr-114 mRNA in nuclei. Right images are longer incubation times of different intestines. (F-H) Characterization of intestinal NHR-114 transgenic protein. (F) Immunostaining of extruded intestines of the EV441 strain (n>20 animals). anti-FLAG detects FLAG::NHR-114 in intestinal cells DAPI visualizes nuclei. Anterior (top); posterior (bottom). antianillin (ANI-2) outlines apical membranes. Scale bar 50 um. (G) Nuclear enrichment of FLAG::NHR-114 in intestinal cells. DAPI-positive bacterial remnants outline intestinal lumen. Scale bar 50 um. (H) Anti-FLAG and anti-tubulin immunoblot. FLAG::NHR-114 protein levels remain unchanged upon nhr-114 RNAi knockdown. Lanes 3 and 4 represent two independent experiments. Effectiveness of RNAi knockdown of endogenous nhr-114 was confirmed by the appearance of steriles in non-transgenic siblings.

Picture from Hwang HY et al. (2002) Proc Natl Acad Sci U S A "The Caenorhabditis elegans vulval morphogenesis gene sqv-4 encodes a ...."

Figure 3. SQV-4 protein expression. (A-H) Selected confocal images of SQV-4 Ab staining in multiple planes of focus along the left-to-right axis of the WT animal. Vulval cells with increased SQV-4 expression are indicated. (A-C) SQV-4 expression is increased in cells containing 10 of 22 vulval nuclei in the early L4 stage. The cells strongly stained with SQV-4 Abs dorsal to the vulval cells are in the uterus. The images are arranged from the left-most plane of focus to the right-most plane. (A) Vulval cells containing four nuclei (P5.ppal, P6.paal, P6.pppl, and P7.papl) on the left side of the worm. (B) Two vulval cells (P5.ppp and P7.paa) in the middle plane of the worm. (C) Vulval cells containing four nuclei (P5.ppar, P6.paar, P6.pppr, and P7.papr) on the right side of the worm. (D-H) SQV-4 expression is increased in cells containing 14 of 22 vulval nuclei in the mid-late L4 stage. The images are arranged from the left-most plane of focus to the right-most plane. (D) Vulval cells containing the same four nuclei as in A. (E) Vulval cells containing two of the four dorsal-most nuclei (P6.papl and P6.ppal), which were not stained at an earlier stage. (F) Vulval cells containing the same two nuclei as in B. (G) Vulval cells containing two of the four dorsal-most nuclei (P6.papr and P6.ppar), which were not stained at an earlier stage. (H) Vulval cells containing the same four nuclei as in C. (I) SQV-4 Abs stain oocytes. A row of oocytes in an adult hermaphrodite stained with SQV-4 Abs is indicated with arrows. The oocyte most proximal to the uterus is located at the lower left. (J-K) SQV-4 protein expression in mutants with abnormal vulval development. (J) Confocal image of a lin-12(gf) mutant fixed and stained with SQV-4 Abs. Each pseudovulva (pv) has three nuclei strongly stained with SQV-4 Abs. The functional vulva (v) has six nuclei strongly stained with SQV-4 Abs. Not all staining is shown here. (K) Confocal image of an L4 lin-11(lf) mutant fixed and stained with SQV-4 Abs. Staining by SQV-4 Abs is limited to one cell in this focal plane. Staining is absent in vulval cells in other focal planes (not shown). (Scale bar = 10 um.)