Picture from Li Q et al. (2018) PLoS Genet "The tumor suppressor BRCA1-BARD1 complex localizes to the synaptonemal complex ...."

Fig 1. GFP::BRC-1 and BRD-1::GFP are expressed in the germ line and respond to stalled/collapsed replication forks. A) GFP fluorescenceof whole worms expressing GFP::BRC-1, BRD-1::GFP, or no GFP (WT). Dashed line denotes germ line with arrows to indicate GFP fluorescence;blue arrows denote GFP signal in embryos; gut granules auto-fluoresce. Scale bar = 100um. B) Schematic of the spatiotemporal organization ofthe hermaphrodite germ line with meiotic stages indicated. C) Proliferating germ cells (premeiotic) expressing GFP::BRC-1 or BRD-1::GFP(green), stained with antibodies against RAD-51 (red), and counterstained with DAPI (blue) in the absence (-HU) and presence of 5uMhydroxyurea (+HU). Blue arrows denote co-localization between GFP and RAD-51. Images are projections through half of the gonad. Scalebar = 5um. D) Pearson's Correlation Coefficient (PCC) measurements between RAD-51 and GFP::BRC-1 or BRD-1::GFP in the absence andpresence of HU. 95% Confidence Intervals are shown. *** p< 0.0001, Mann-Whitney test; n = 10 nuclei.

Picture from Olsen A et al. (2006) Science "Checkpoint proteins control survival of the postmitotic cells in ...."

Figure 1. CID-1 regulates stress resistance. (A) Longitudinal thermotolerance assay of cid-1(rf34::Tc4) mutant lines. (+/+) indicates wild-type cid-1 gene. (B) Longitudinal thermotolerance assay of rrf-3(pk1426) cid-1(RNAi) worms. The rrf-3(pk1426) mutation renders worms more sensitive to RNAi. The complete thermotolerance data can be found in table S1. (C) HU-resistance assay. (Left) Wild-type N2 worms showed arrested development at the L1/L2 stage when eggs were placed on HU plates. (Right) cid-1(rf34::Tc4) mutants develop into fertile adults at the same concentration of HU. (D) Life span of rrf-3(pk1426) cid-1(RNAi) worms at 20C. (E) Transgenic worms expressing a CID-1::GFP gene fusion. (Top) Composite fluorescent and bright-field micrographs and corresponding fluorescent micrographs of the fusion protein localized to the first layer of intestinal cells (Int 1) and to posterior intestinal cells. (Bottom) A high-resolution fluorescent micrograph of the fusion protein localized to the cytoplasm of the posterior intestinal cells.

Picture from Dammermann A et al. (2008) J Cell Biol "SAS-4 is recruited to a dynamic structure in newly forming centrioles that is ...."

Figure 2. Kinetic profiles for SAS-6 recruitment in control, sas-5(RNAi), sas-4(RNAi), and HU-treated embryos. (A) High-resolution images of GFP:SAS-6 recruited to RFP-labeled sperm centrioles in embryos generated using the mating scheme in Fig. 1 A. Times (in seconds relative to cytokinesis onset) correspond to meiosis (-1,470 to -1,370 s), early (-1,180 to -1,156 s) and mid (-1,005 to -972 s) S phase, and late prophase (-281 to -267 s). (B) Normalized individual measurements of the integrated GFP:SAS-6 intensity coincident with sperm centriolar RFP signal (see Materials and methods for details on normalization). (C) Kinetic profiles for the recruitment of GFP:SAS-6 in control and sas-5(RNAi) embryos. Data points are the mean of the normalized GFP intensity measurements collected during the 200-s interval centered on that point. (D) Depletion of SAS-4 does not affect the kinetics of centriolar GFP:SAS-6 recruitment or loss. (E) Recruitment profile of centriolar GFP:SAS-6 in embryos treated with 75 mM HU. The profile was generated as in C and D except that sequences were time-aligned with respect to centriole separation, which occurs at the same time after meiosis II anaphase in control and HU-treated embryos (Videos 1 and 2, available at http://www.jcb.org/cgi/content/full/jcb.200709102/DC1). The control dataset plotted here is the same as in B-D except that the means were recalculated for the indicated 200-s intervals after converting seconds before cytokinesis onset to seconds after centriole separation. All error bars indicate the 90% confidence interval.

Picture from AlHarbi S et al. (2023) MicroPubl Biol "Characterizing a standardized BioPart for PVQ-specific expression in C. ...."

Figure 1. Expression and Characterization of Pnlp-17 in PVQ Neurons of C. elegans:A. Pnlp-17::mScarlet::tbb-2 3'UTR expression is specific to PVQ neurons from extrachromosomal arrays in hermaphrodites (top panel) and males (bottom panel). B. Single-copy Pnlp-17::mScarlet transgenes inserted by MosTI (El Mouridi et al., 2022) are easily visible in hermaphrodites. 20x magnification. Scale bars: 100 µm. C. Characteristics and quantification of the short 300 bp nlp-17 promoter. D. Schematic of standard BioPart expression vectors for PVQ expression. Unmodified gfp or mScarlet vectors can be used to identify PVQ neurons. Standard restriction sites allow cell-specific expression with N-terminal (ApaI) and C-terminal (SmaI) fluorophore fusions or complete fluorophore exchange (BsaI). The vectors are compatible with single-copy insertions (MosTI) and expression from extra-chromosomal arrays. E. The short 300 bp promoter is easily synthesized and has been included as a standard feature in an online tool for optimizing transgene design and synthesis (www.wormbuilder.org/transgenebuilder) (Vargas-Velazquez, El Mouridi, Alkhaldi, and Frøkjær-Jensen C, manuscript in preparation).

Picture from Gengyo-Ando K et al. (2024) MicroPubl Biol "Domain 3a mutation of VPS33A suppresses larval arrest phenotype in the loss of ...."

Figure 1. vps-331 mutations suppress vps-45(0) ts larval arrest and lethality:(A) Genomic region of the vps-33.1 gene. Exons are indicated by gray boxes, mutation sites in jq28 and syb8647 are underlined, and amino acid substitutions are indicated below. The VPS-33.1 proteins contain three domains, with domain 3 split into domain 3a and domain 3b. The suppressor mutation associated residue methionine 376 (M376I, arrowhead) is located in the domain 3a. The amino acid sequence alignments of VPS33A and VPS45 are shown below [VPS33A: Homo sapiens (NP_075067.2), Danio rerio (NP_001093443.1), Drosophila melanogaster (AAF48972), Caenorhabditis elegans (CCD61712.1), Saccharomyces cerevisiae (KZV09647.1), VPS45: Homo sapiens (NP_009190), Danio rerio (NP_001243585.1), Drosophila melanogaster (AAF54403.1), Caenorhabditis elegans (NP_741714.1), Saccharomyces cerevisiae (CAA96801.1)]. The conserved amino acids of the VPS33 or VPS45 proteins are highlighted in grey and the conserved amino acids of both proteins are highlighted in light blue. The M376 is indicated by a red box. The strictly conserved proline (proline 363 in C. elegans) in the domain 3a (Hu et al., 2011) is indicated by a blue asterisk. (B) Structure of the C. elegans VPS-33.1 protein based on I-TASSER structural modeling. The 3D model with the highest confidence score is shown. The distal tip of domain 3a which is expected to interact with SNAREs/SNARE complex (Pieren, Schmidt and Mayer, 2010; Hu et al., 2011; Baker, Jeffrey and Hughson, 2013) is shown in green. The residue M376 is indicated by a space-filling representation in red. The conserved proline, P363, is highlighted in blue. (C) Representative images of wild-type, vps-45(0), vps-33.1(jq28);vps-45(0), vps-33.1(syb8647);vps-45(0), vps-33.1(syb8647) animals grown from eggs at 25°C for 76 h. vps-45(0) represents the null allele vps-45(tm246). Scale bars, 200 µm. (D) Quantification of body length of animals in (C) (n=23-29). mean±SD. Statistics: one-way ANOVA with Dunnet's post hoc test. ****p<0.0001. ns: not significant. (E) Percentage of adults in the animals measured in (D) (n=23-29). ***p<0.001. ns: not significant. Statistics: Fisher's exact test, two-tailed.

Picture from Lu Y et al. (2014) PLoS One "Centrosome/Cell cycle uncoupling and elimination in the endoreduplicating ...."

(B-F) and (G-I) SPD-2 and SAS-4 foci are detectable up to the L1/L2 transition. The signal is no longer detectable by the L3 stage. The nuclear localized SPD-2 is absent following spd-2(RNAi) in (F). Animals were stained with DAPI (red) and antiSPD-2 (green) in (B-F) or anti-SAS-4 (green) in (G-I). Arrowheads indicate centrioles, while asterisks indicate intestinal nuclei. The insets represent magnified SPD-2 or SAS-4 signal of the highlighted regions (white rectangles). The circles in (E) and (I) highlight the germ cells with either SPD-2 foci or SAS-4 foci. (J) Quantification of centriole numbers and described in (B-E) and (G-I) based on SPD-2 or SAS-4 detection. n=56 for each stage. (K) Centriole duplication is uncoupled from the endocycles in the lateral hypodermal V cells in the late L1 stage. The centriole appears to be uncoupled from S-phase in the anterior endoreduplicating daughter cell but remains coupled to DNA replication in the mitotic posterior daughter. The square brackets indicate the daughter cells from a common mother V cell, while arrowheads indicate the centrioles. The inset is a magnified view of the region delineated by the white rectangle. A, anterior; P, posterior. hyp7 marks the hyp7 nucleus. (L and M) Supernumerary centrioles are detected in HU-treated L1 animals (M) but not in the control (L). The inset is a magnified SPD-2 signal of the region delineated by the white rectangle. Arrowheads indicate centrioles, while asterisks indicate intestinal nuclei. (N and O) Centriole duplication occurs once in response to S phase, but centrioles do not overduplicate and are not eliminated during un-quantized DNA re-replication. Heterozygous in (N) and homozygous cul-4 (gk434) mutants in (O) were stained with DAPI (red) and SPD-2 (green) respectively. The insets show the SPD-2 signal in the regions outlined by the white rectangles. Arrowheads indicate centrioles. Scale bar, 5 um.doi:10.1371/journal.pone.0110958.g002

Picture from Lu Y et al. (2014) PLoS One "Centrosome/Cell cycle uncoupling and elimination in the endoreduplicating ...."

Figure 2. Centrioles no longer duplicate in endocycling cells prior to their elimination. (A) SPD-2::GFP signal can be seen throughout the germline until oogenesis in (a), while in (b) it is undetectable in the intestinal cells in adult hermaphrodites. a9 and b9 are high magnification images of the GFP signal in the field identified by the two white rectangles a and b. (B-F) and (G-I) SPD-2 and SAS-4 foci are detectable up to the L1/L2 transition. The signal is no longer detectable by the L3 stage. The nuclear localized SPD-2 is absent following spd-2(RNAi) in (F). Animals were stained with DAPI (red) and antiSPD-2 (green) in (B-F) or anti-SAS-4 (green) in (G-I). Arrowheads indicate centrioles, while asterisks indicate intestinal nuclei. The insets represent magnified SPD-2 or SAS-4 signal of the highlighted regions (white rectangles). The circles in (E) and (I) highlight the germ cells with either SPD-2 foci or SAS-4 foci. (J) Quantification of centriole numbers and described in (B-E) and (G-I) based on SPD-2 or SAS-4 detection. n$56 for each stage. (K) Centriole duplication is uncoupled from the endocycles in the lateral hypodermal V cells in the late L1 stage. The centriole appears to be uncoupled from S-phase in the anterior endoreduplicating daughter cell but remains coupled to DNA replication in the mitotic posterior daughter. The square brackets indicate the daughter cells from a common mother V cell, while arrowheads indicate the centrioles. The inset is a magnified view of the region delineated by the white rectangle. A, anterior; P, posterior. hyp7 marks the hyp7 nucleus. (L and M) Supernumerary centrioles are detected in HU-treated L1 animals (M) but not in the control (L). The inset is a magnified SPD-2 signal of the region delineated by the white rectangle. Arrowheads indicate centrioles, while asterisks indicate intestinal nuclei. (N and O) Centriole duplication occurs once in response to S phase, but centrioles do not overduplicate and are not eliminated during un-quantized DNA re-replication. Heterozygous in (N) and homozygous cul-4 (gk434) mutants in (O) were stained with DAPI (red) and SPD-2 (green) respectively. The insets show the SPD-2 signal in the regions outlined by the white rectangles. Arrowheads indicate centrioles. Scale bar, 5 um.doi:10.1371/journal.pone.0110958.g002

Picture from Hu J et al. (2007) Mol Biol Cell "STAM and Hrs down-regulate ciliary TRP receptors."

Figure 2. STAM-1A coexpresses with LOV-1 and PKD-2 in male-specific neurons and shows an endosome-like distribution pattern. (A) Anatomy of C. elegans male and PKD-2-expressing neurons. Modified from (Hu et al., 2006). Top, differential interference contrast (DIC) side view image of an adult C. elegans male oriented head (left) to ventral up tail (right). Middle, lov-1 and pkd-2 are expressed in male head CEM neurons (left) and tail ray B type neurons (except R6B) and the hook HOB neuron (right). Bottom, confocal micrographs of PKD-2::GFP expression pattern in male head and tail. The cilium, dendrite, cell body, and axon of the CEMD (dorsal CEM) and CEMV (ventral CEM) neuron are drawn in the male head diagram. CEM cilia are in the nose region (dashed rectangular box). The pharynx is green. Positions of nuclei of all lov-1- and pkd-2-expressing cells in the C. elegans adult male tail (modified from Sulston et al., 1980). Ray neurons are arranged as nine left-right bilateral pairs, numbered 1-9, anterior to posterior (ventral up view shown here). Rays are required for response to contact with a potential mate (Liu and Sternberg, 1995). HOB is an asymmetric ciliated hook neuron that mediates vulva location behavior (Liu and Sternberg, 1995). For simplicity, only the dendrite of R3B left and HOB is shown. The R3B cilium is indicated by a dashed rectangular box. (B) Confocal micrographs of Pstam-1::DsRed2 (using the stam-1 promoter to drive DsRed2 expression) and PKD-2::GFP double-labeled C. elegans males. In all figures, adult males are oriented anterior to posterior, with the head pointing left and the tail pointing right. stam-1 coexpresses with pkd-2 in CEMs (head), RnBs, and HOB (tail, arrow points to HOB neuron). (C) With its native promoter, STAM-1A::GFP displays an endosomal-like pattern throughout the neuronal cell bodies. Note that STAM-1A also localizes to the cilia base of cilia in enlarged figure (right panel). In this and all subsequent figures, the arrow marks the ciliary base, and the arrowhead marks the approximate end of the cilium. (D) To clearly visualize the localization pattern of STAM-1A in polycystin-expressing neurons, the pkd-2 promoter is used to drive stam-1a cDNA expression only in CEMs, RnBs, and HOB neurons. STAM-1A localizes to the ciliary base (arrow), but is excluded from the cilium proper (region between arrow and arrowhead). Large green puncta are observed along distal dendrites of RnB ray neurons. Enlarged figures show the cilia region of CEM and R3B. Scale bars in rightmost C and D panels, 5 um; in all other micrographs scale bars, 10 um.