Picture from Lee MH et al. (2007) PLoS Genet "Conserved regulation of MAP kinase expression by PUF RNA-binding ...."

Immunocytochemistry of dissected adult hermaphrodite germlines stained using both MPK-1 antibodies (G) and DAPI (H). Distal end, arrowhead; dotted lines, boundaries between regions of germline maturation [MR (mitotic region), TZ (transition zone), PR (pachytene region), OO (oocytes), SP (sperm)].

Picture from Lee MH et al. (2007) PLoS Genet "Conserved regulation of MAP kinase expression by PUF RNA-binding ...."

In situ analysis of dissected adult hermaphrodite germlines. (D) Total mpk-1 RNA was assessed using the mpk-1ab antisense probe shown in (A). (E) mpk-1b RNA was assessed using an isoform-specific antisense probe shown in (A). (F) Negative control, using an mpk-1b-specific sense probe. [MR (mitotic region), TZ (transition zone), PR (pachytene region), OO (oocytes), SP (sperm)].

Picture from Lamont LB et al. (2004) Dev Cell "FBF-1 and FBF-2 regulate the size of the mitotic region in the C. elegans ...."

Figure 4. FBF-1 and FBF-2 Proteins Are Expressed in Distinct Patterns that Are Responsive to FBF Activity. (A, B, E-G) Extruded adult germlines, distal end is to left. Germlines were treated identically, and confocal images were taken with the same settings at the same magnification for comparison of (A) to (B) and of (E) and (F) to (G). White arrowheads highlight the distal and proximal boundaries of staining.(A) FBF-1 staining in wild-type germline.(B) FBF-1 increases in both extent and amount in fbf-2(q738) germlines.(C and D) Replicate gels run with recombinant proteins: Coomassie-stained polyacrylamide gel (C) and Western blot with anti-FBF-2-specific antibodies (D).(E) FBF-2 staining in wild-type germline.(F) FBF-2 staining in fbf-2(q738) single mutant.(G) FBF-2 increases in amount in fbf-1(ok91) single mutant germ lines, with respect to wild-type.(H) Summary of effects of fbf-1 and fbf-2 mutations on mitotic region. MR, mitotic region; n.d., no data. Distance from DTC scored in nuclei for MR length and proximal end of FBF staining. Asterisk indicates value is significantly different from wild-type (p < 0.001).

Picture from Park YS et al. (2000) Biochem Biophys Res Commun "Alternative splicing of gar-1, a Caenorhabditis elegans G-protein-linked ...."

Figure 5. RT-PCR of the three gar-1 mRNAs at various develop- mental stages. RT-PCR was performed, with a primer pair flanking the i3 loop (MF-i3/MR-i3), using stage-specific RNA as a template (see Materials and Methods). The PCR products were analyzed on an 1% agarose gel. The expected sizes of the PCR products for gar-1a, gar-1b, and gar-1c are 1281, 1188, and 1008 bp, respectively (from top to bottom). E, embryo; L1, first larval stage; L2, second larval stage; L3, third larval stage; L4, fourth larval stage; and A, young adult.

Picture from Lee MH et al. (2007) PLoS Genet "Conserved regulation of MAP kinase expression by PUF RNA-binding ...."

Figure 1. mpk-1 Expression in the C. elegans Germline (A) Schematics of mpk-1a and mpk-1b mRNAs. Box, exon; connecting line, intron; ATG, initiation codon; TAG, termination codon. Below schematics: thick bars, extent of probes used for in situ hybridization; arrows, primer pairs used for RT-PCR. (B) Semiquantitative RT-PCR of RNA prepared from adult hermaphrodites that either had an essentially normal germline [glp-1(q224) grown at 15 C], or had virtually no germline [glp-1(q224) grown at 25 C]. unc-54 was used as a control. (C) Western blot. MPK-1A protein is 45 kDa, MPK-1B is 55 kDa, and alfa-TUB is alfa-tubulin. Proteins were extracted from adult hermaphrodites that were either wild-type (wt), glp-1(q224) grown at 25 C (GL-), or mpk-1(ga117) putative null homozygotes [mpk-1(0)]. (D-F) In situ analysis of dissected adult hermaphrodite germlines. (D) Total mpk-1 RNA was assessed using the mpk-1ab antisense probe shown in (A). (E) mpk-1b RNA was assessed using an isoform-specific antisense probe shown in (A). (F) Negative control, using an mpk-1b-specific sense probe. (G-L) Immunocytochemistry of dissected adult hermaphrodite germlines. All were stained using both MPK-1 antibodies (G, I, K) and DAPI (H, J, L). Distal end, arrowhead; dotted lines, boundaries between regions of germline maturation [MR (mitotic region), TZ (transition zone), PR (pachytene region), OO (oocytes), SP (sperm)]; PEX (pachytene exit defect). (G, H) Same wild-type germline. (I, J) Same mpk-1(0) germline. (K, L) Same mpk-1b(RNAi) germline.

Picture from Thompson BE et al. (2005) Development "Dose-dependent control of proliferation and sperm specification by ...."

Figure 6. FBF represses fog-1 expression in vivo. (A-J) Arrow indicates distal end. Scale bars: 10 um. (A-E,G-J) Animals or dissected germlines stained with anti-FOG-1 antibodies. Broken lines indicate germlines in A-D. (A) Wild-type L2. FOG-1 is barely detectable. (B) Wild-type L3. FOG-1 is abundant in proximal germline where cells are destined to become sperm; FOG-1 is low in distal germline where cells remain in mitotic cell cycle. (C,D) L2 fbf-1 fbf-2. FOG-1 is more abundant than in wild type (compare with A) and its distribution is not graded. (D) L3 fbf-1 fbf-2. FOG-1 is more abundant than in wild type (compare with B) and its distribution is not graded. (E) Wild-type adult male germline. MR, mitotic region; TZ, transition zone. FOG-1 levels are low in mitotic region and become high as germ cells begin entry into meiosis in proximal part of mitotic region and transition zone. FOG-1 disappears in more mature stages of meiotic prophase. (F) Wild-type adult male germline hybridized with fog-1 antisense probe. fog-1 mRNA has same pattern as FOG-1 protein (compare with E). Sense probe has no detectable staining (not shown). (G) fog-1(q250) homozygous male germline. No FOG-1 protein is detectable. (H) Wild-type adult hermaphrodite germline. No FOG-1 protein is detectable. (I) Adult gld-3 nos-3 tumorous germline. FOG-1 levels are low. (J) Adult fbf-1 fbf-2 gld-3 nos-3 tumorous germline. FOG-1 levels increase in the absence of FBF (compare with I).