Picture from Peel N et al. (2017) PLoS Genet "Protein Phosphatase 1 Down Regulates ZYG-1 Levels to Limit Centriole ...."

S3 Fig. Localization of I-2^SZY-2 , PP1β^GSP-1 and SDS-22. GFP fusions of the indicated protein were expressed in the embryo. I-2^SZY-2 Is enriched in the nuclei. Pronuclei meeting during the first cell cycle is shown. PP1β^GSP-1 is enriched on the chromatin throughout mitosis. Anaphase of the first cell cycle is shown. SDS-22 is weakly localized to the spindle. Metaphase of the first cell cycle is shown.

Picture from Crawford MW et al. (2023) MicroPubl Biol "Mutations in the NXF-1:NXT-1 mRNA export complex affect gene-expression driven ...."

Figure 1. Missense mutations in nxf-1 and nxt-1 partially suppress Phsp-16.41::peel-1 and Phsp-16.41::eGFP::his-44 but not endogenous peel-1:(A) Schematic of NXF-1 and NXT-1 indicating functional domains (see text for details) and the mutations that were identified in our screen. (B-C) Quantification of Phsp-16.41::peel-1 lethality as the fraction of moving (living) worms at different times after a 2 hr heat shock at 34°C; n=3-5, each replicate represents 50 animals; error bars represent standard error of the mean. (D) Schematic of expected outcomes and quantification of dead progeny from animals heterozygous for the zeel-1(tm3419) deletion and homozygous for nxf-1(yak65) with endogenously expressed peel-1. WT control shows quantification of dead progeny from animals heterozygous for zeel-1(tm3419). n=7-9, each n represents 12-127 animals; ns - not significant (p>0.05); error bars represent standard deviation. (E) Relative fluorescence of Phsp-16.41::GFP in an nxf-1(yak65) mutant normalized to control; n=3 replicates, each replicate group represents 8-17 animals; bars represent means. Statistics: unpaired two-tailed t tests.

Picture from Seidel HS et al. (2011) PLoS Biol "A novel sperm-delivered toxin causes late-stage embryo lethality and ...."

Figure 2. peel-1 is expressed exclusively in sperm and carries an N-terminal sperm localization signal. (A) Adult male expressing Ppeel-1::GFP. The tube-like gonad begins at the asterisk, extends toward the head, folds over on itself, and then extends toward the tail. The arrow and arrowhead indicate spermatocytes and sperm, respectively. Fluorescence outside the gonad is auto-fluorescence. Nomarski and fluorescence channels are overlaid. (B) Secondary spermatocytes, budding spermatids, and mature spermatids expressing either untagged GFP or GFP tagged with the N-terminal 12 amino acids of PEEL- 1 (PEEL-112a.a.::GFP). Arrows indicate residual bodies. Nomarski and fluorescence channels are overlaid. (C) Relative expression levels of peel- 1 and spe-9 in him-5(e1490) and him-5(e1490) fem-1(hc17ts) adult males at the permissive (15uC) and restrictive (25uC) temperatures. him- 5(e1490) was included to aid in collection of males. Expression levels were calculated relative to the him-5(e1490) fem-1(hc17ts) 15uC sample. Runs were performed in triplicate and standard deviations are shown. * p,1025, one-tailed Student's t test on the normalized expression levels. n/s, p.0.05.

Picture from Seidel HS et al. (2011) PLoS Biol "A novel sperm-delivered toxin causes late-stage embryo lethality and ...."

Figure 3. PEEL-1 localizes to fibrous body-membranous organelles. (A) Diagram of spermatogenesis and fibrous body-membranous organelle (FB-MO) development, adapted with permission from [16]. FB-MOs develop in pachytene spermatocytes as membrane-bound organelles having a head region separated by a collar-like constriction from a set of membrane folds (lower panel; red). As spermatogenesis proceeds, the membrane folds grow and extend into arm-like protrusions, enveloping bundles of polymerized Major Sperm Protein, referred to as fibrous bodies (hatched region). Coincident with budding of spermatids from the residual body, the membrane folds of FB-MOs retract, and the fibrous bodies depolymerize into the cytoplasm. The FB-free MOs then move to a position just inside the plasma membrane, and upon sperm activation, they fuse with the plasma membrane opposite the pseudopod. (B-F) Nomarski and fluorescence images of spermatocytes and sperm expressing PEEL-1::GFP. Panels in (B) show the proximal arm of a male gonad, oriented with pachytene spermatocytes towards the left (bracketed region). Arrow and arrowhead indicate primary and secondary spermatocytes, respectively. Panels in (C-F) show higher resolution images of the following stages: secondary spermatocyte (C), budding spermatids (D), unactivated spermatid (E), and activated spermatozoan (F). (G-K) Spermatids were dissected from peel-1(+) (G-I) or peel-1(D) (J-K) males and stained with anti-PEEL-1 (green) and the FB-MO marker, 1CB4 (red) [64]. Nuclei are stained with DAPI (blue).

Picture from Aviles S et al. (2024) MicroPubl Biol "New alleles of nlp-2 , nlp-22 , and nlp-23 demonstrate ...."

Figure 1. The RPamide neuropeptides are dispensable for stress-induced sleep:(a) Gene structures and alleles for the RPamide neuropeptides. (b) Average minutes of movement quiescence in 10-minute windows during L4 developmentally-timed sleep in wild-type (N=31) and nlp-22(stj313) (N=33) animals, wild-type (N=35) and nlp-23(stj310) (N=36) animals, and wild-type (N=26), nlp-22(stj313) (N=30), and nlp-22(stj313);nlp-23(stj311) (N=60) animals. (c) Total minutes of movement quiescence during L4 developmentally-timed sleep in wild-type (N=31) and nlp-22(stj313) (N=33) animals, wild-type (N=35) and nlp-23(stj310) (N=36) animals, wild-type (N=26), nlp-22(stj313) (N=30), and nlp-22(stj313);nlp-23(stj311) (N=60) animals, and wild-type (N=38), nlp-22(stj313);nlp-23(stj311) (N=34), and nlp-2(stj351);nlp-22(stj313);nlp-23(stj311) (N=82) animals. (d) Total minutes of movement quiescence during UV-induced sleep in wild-type (N=33) and nlp-22(stj313) (N=31) animals, wild-type (N=44) and nlp-23(stj310) (N=49) animals, wild-type (N=55), nlp-22(stj313);nlp-23(stj311) (N=41), and nlp-2(stj351);nlp-22(stj313);nlp-23(stj311) (N=50) animals. For both (c) and (d) statistical significance was calculated by Student's t-test (2 genotypes) or one-way ANOVA followed by Tukey's test (3 genotypes)(*p<0.05, **p<0.01, ***p<0.001).

Picture from Revathi, Kandan et al. MicroPubl Biol

Figure 1. Expression of peel-1 with its endogenous introns kills all array-positive animals:(A) Cartoon diagram of the peel-1 expression cassette. 1. hsp-16.41 promoter, 2. hsp-16.41 5' UTR, 3&#x2012;6. peel-1 exons, and 7. tbb-2 3' UTR. The restriction enzymes used are indicated above, with their respective positions in parentheses. The blue lines between exons represent the introns. Note: Intron-1 was not included; instead, the last 16 nucleotides of exon-1, which contains the last six nucleotides of peel-1 5' UTR and the first 10 nucleotides of the coding region, were included in the PCR primer used for amplifying the rest of peel-1 coding region. (B) Metrics for the four microinjection experiments. Names of the transgenes are given below the corresponding bars. (C) Results of PCR amplification for the 55 animals tested for pKS250. In this experiment, the left integration junction was amplified using primers KS6455 and KS6457. The expected 1.8-kb band is indicated by the arrow on the left. The second band about 0.9 kb seen in all lanes is due to non-specific amplification. M - DNA molecular weight markers. (D) From plates represented by the grey bars in (B), wildtype (rescued for the unc-119 mutation) animals were cloned. All such animals in each plate were assumed to be the descendants of a single animal with successful transgene integration. Based on this, each plate was assumed to represent one independent line. The number of wildtype animals recovered and cloned from each line is mentioned above the corresponding bar. In some plates (Line-2, Line-6, and Line-16), only one wildtype animal was found to have survived the heat-shock. After collecting a few embryos, the cloned animals were subjected to PCR using integration-specific primers to determine the presence of the integrated transgene. (E) A representative image showing transgene expression in the germ line.

Picture from Tian Y et al. (2010) Cell "C. elegans screen identifies autophagy genes specific to multicellular ...."

(M-O) The epg-5 reporter is expressed ubiquitously in embryos (N) and widely in larvae (O). EPG-5 is cytoplasmic. (M) Nomarski image of the embryo in N.

Picture from Zhao G et al. (2007) Genome Res "Identification of muscle-specific regulatory modules in Caenorhabditis ...."

(N) K10G6.3::gfp expression in the pharyngeal muscle (arrow) and neurons.

Picture from Ou G et al. (2007) Mol Biol Cell "Sensory ciliogenesis in Caenorhabditis elegans: assignment of IFT components ...."

Schematics and fluorescence microscopy of C. elegans neuronal cilia. (J) Cilia structure schematics adapted from Ward et al. (1975). (N) Fluorescence microscopic images of neuronal cilia. (J and N) The 'fork' shaped AWB cilia, which are visualized using a GFP transcriptional fusion of a seven-span transmembrane receptor, STR-1 (N).

Picture from Vasudevan A et al. (2024) MicroPubl Biol "Physical presence of chemical synapses is necessary for turning behavior of ...."

Figure 1. pre-SVs do not turn into the synaptic branch upon loss of connection to chemical synapses in the C. elegans PLM neuron: For all plots shown in the figure, individual data points are represented as solid circles. The summary statistics are depicted using a boxplot with whiskers. Upper whisker = minimum(maximum value in the distribution, Q3 + 1.5 * IQR) and lower whisker = maximum(minimum value in the distribution, Q1 - 1.5*IQR), where IQR is inter-quartile range.A) Schematic of the C. elegans PLM neuron, representing the laser ablation paradigm for the 'synapse cut'. The red cross marks the site at which laser ablation is performed, such that the connection between the chemical synapses and the synaptic branch is removed. Proportion of 'turning', 'pausing', and 'straight' vesicles is plotted as a percentage of total anterograde pre-SVs that reach the branch point in wild type- jsIs821 animals. N(before cut)=10 animals, n(before cut- 'turning')=103 vesicles, n(before cut-'pausing')= 90 vesicles, n(before cut-'straight')= 20 vesicles. N(after cut) =10 animals, n(after cut-'turning')=6 vesicles, n(after cut-'pausing')= 121 vesicles, n(after cut-'straight')= 15 vesicles. Wilcoxon rank sum test with continuity correction conducted to test for statistical significance, ****p<0.0001. All comparisons are made to the 'before cut' category.B) Schematic of PLM neuron, representing the laser ablation paradigm for the 'control cut'. The red cross marks the site at which laser ablation is performed along the distal process. This site is chosen such that its distance from the branch point is roughly equal to the length of the PLM synaptic branch (~30µm). Proportion of 'turning', 'pausing', and 'straight' vesicles is plotted as a percentage of total anterograde pre-SVs that reach the branch point in wild type- jsIs821 animals. N(before cut)=9 animals, n(before cut- 'turning')=95 vesicles, n(before cut-'pausing')=87 vesicles, n(before cut-'straight')=19 vesicles. N(after cut) =9 animals, n(after cut-'turning')=101 vesicles, n(after cut-'pausing')=95 vesicles, n(after cut-'straight')=15 vesicles. Wilcoxon rank sum test with continuity correction conducted to test for statistical significance. All comparisons are made to the 'before cut' category and are not statistically significant.C) Schematic of the PLM neuron depicting the different categories of pre-SVs transported at the branch point, namely, i) anterograde pre-SVs transported from the cell body to the branch point, ii) retrograde pre-SVs transported from the synaptic branch to the branch point, and iii) retrograde pre-SVs transported from the distal process to the branch point.D) Total flux of pre-SVs reaching the branch point in the anterograde and retrograde directions over a period of 5mins in the 'synapse cut' paradigm. N(before cut)=10 animals, n(before cut- 'anterograde')=213 vesicles, n(before cut- 'retrograde from branch')=26 vesicles, n(after cut- 'anterograde')=142 vesicles, n(after cut-'retrograde from branch')=19 vesicles. Wilcoxon rank sum test with continuity correction conducted to test for statistical significance, all comparisons are non-significant. All comparisons are made to the 'before cut' category.E) Total flux of pre-SVs reaching the branch point in the anterograde and retrograde directions over a period of 5mins in the 'control cut' paradigm. N(before cut)=9 animals, n(before cut- 'anterograde')=201 vesicles, n(before cut- 'retrograde from distal process')=119 vesicles, n(after cut- 'anterograde')=211 vesicles, n(after cut-'retrograde from branch')=72 vesicles. Wilcoxon rank sum test with continuity correction conducted to test for statistical significance, all comparisons are non-significant. All comparisons are made to the 'before cut' category.F) Schematic of the PLM branch point depicting the mobilization of pre-SVs from the paused vesicle cluster at the branch point into different compartments of the neuron.G) Total flux of pre-SVs mobilizing from the paused vesicle cluster at the branch point over a period of 5mins in the 'synapse cut' paradigm. N(before cut)=10 animals, n(before cut-'Into branch')=50 vesicles, n(before cut-'Into distal process')=46 vesicles, n(before cut-'Towards cell body')=24 vesicles. N(after cut) =10 animals, n(after cut-' Into branch')=45 vesicles, n(after cut-'Into distal process')=39 vesicles, n(after cut-'Towards cell body')=21 vesicles. Wilcoxon rank sum test with continuity correction conducted to test for statistical significance, all comparisons are non-significant. All comparisons are made to the 'before cut' category.H) Total flux of pre-SVs mobilising from the paused vesicle cluster at the branch point over a period of 5mins in the 'control cut' paradigm. N(before cut)=9 animals, n(before cut-'Into branch')=45 vesicles, n(before cut-'Into distal process')=46 vesicles, n(before cut-'Towards cell body')=20 vesicles. N(after cut) =9 animals, n(after cut-'Into branch')=42 vesicles, n(after cut-'Into distal process')=41 vesicles, n(after cut-'Towards cell body')=22 vesicles. Wilcoxon rank sum test with continuity correction conducted to test for statistical significance, all comparisons are non-significant. All comparisons are made to the 'before cut' category.