Picture from Power KM et al. (2024) MicroPubl Biol "osm-5p- driven fluorophores are differentially expressed in ...."

Figure 1. osm-5p-driven soluble GFP differs in brightness in ccpp-1Δ and nekl-4Δ mutant ciliated neurons: a. Widefield images of soluble osm-5p::GFP in the phasmid soma, sum intensity projections. Scale bar = 5 µm. b. Quantification of mean fluorescence intensity and integrated density of osm-5p::GFP fluorescence in the phasmid soma. au = arbitrary unit. Mean ± SEM; * indicates p ≤ 0.05, ** indicates p ≤ 0.01, *** indicates p ≤ 0.001, **** indicates p ≤ 0.0001 by Kruskal-Wallis one-way ANOVA with post hoc Dunn's correction for multiple comparisons. n is indicated above each genotype. Black indicates significance relative to WT, red indicates significance relative to ccpp-1Δ.

Picture from Chen Z et al. (2013) Neuron "Two insulin-like peptides antagonistically regulate aversive olfactory ...."

Figure 5. ins-6 Negatively Regulates ins-7 Expression in URX Neurons (A) Representative images for the expression of yxIs13[Pins-7::gfp] reporter in URX neurons in wild-type animals and ins-6(tm2416) mutants. Red fluorescence in URX is generated by the expression of the Pgcy-36::mcherry transgene. (B and C) The expression of yxIs13[Pins-7::gfp] in URX neurons is increased in ins-6(tm2416) mutants (B) and this defect is rescued by the wild-type ins-6 genomic DNA (Pins-6::ins-6) (C). Each circle indicates a measurement of yxIs13[Pins-7::gfp] fluorescence intensity of the specified neuron from one animal. Student's t test; **p < 0.01; mean +/- SEM; AU, artificial unit. (D and E) The expression of yxIs13[Pins-7::gfp] in ASK (D) and ASI (E) neurons was comparable between wild-type animals and ins-6(tm2416) mutants. Each circle indicates a measurement of yxIs13[Pins-7::gfp] fluorescence intensity of the specified neuron from one animal. Student's t test; n.s., not significant; mean +/- SEM. AU, artificial unit. (F) Expression of ins-6 in URX using the gcy-36 promoter rescued the learning defect in ins- 6(tm2416) mutants. The learning ability of transgenic animals was compared with that of nontransgenic siblings using the paired Student's t test. *p < 0.05; n R 5 assays; mean +/- SEM. (G) Increasing URX expression of ins-7 (Pgcy- 36::ins-7) disrupts learning in wild-type animals. The learning ability of transgenic animals was compared with that of nontransgenic siblings using the paired Student's t test. *p < 0.05; nR5 assays; mean +/- SEM.

Picture from Chen Z et al. (2013) Neuron "Two insulin-like peptides antagonistically regulate aversive olfactory ...."

Figure 8. INS-6 and INS-7 Have Distinct Signal Properties that Respond to Environmental Cues (A and B) Expression of ins-7 in the ASI neurons (Pstr-3::ins-7) does not rescue the learning defect of ins-6(tm2416) mutants (A); conversely, expression of ins-6 in URX (Pgcy-36::ins-6) does not rescue the learning phenotype of ins-7(tm2001) in ins-6(tm2416); ins-7(tm2001) double mutants (B). Paired Student's t test; n.s., not significant, n R 5 assays, mean ± SEM. (C and D) Representative images (C) and quantification (D) of yxIs13[Pins- 7::gfp] expression in URX in dauers and age-matched non-dauers. In (D), each circle indicates a measurement of yxIs13[Pins-7::gfp] expression in URX from one animal. Student's t test; ***p < 0.001; mean ± SEM; AU, artificial unit.

Picture from Lohmer LL et al. (2016) PLoS Genet "A Sensitized Screen for Genes Promoting Invadopodia Function In Vivo: CDC-42 and ...."

S2 Fig. gdi-1 is upregulated in the AC throughout invasion. (A) A transcriptional reporter (gdi-1>GFP) revealed gdi-1 expression in the AC and vulval precursor cells throughout invasion. Expression of gdi-1 is specifically upregulated in the AC. (B) An AC-specific full-length fusion of GFP to GDI-1 (cdh3>GFP::gdi-1) showed cytosolic distribution with no specific subcellular enrichment. (C, D) The gdi-1 RNAi construct reduced levels of an AC expressed GDI1 reporter (cdh3>GFP::gdi-1; n = 15 wild type and 14 gdi-1 RNAi treated animals; p < 0.0001, Student's t-test). (E, F) The gdi-1 RNAi construct from C. elegans did not alter the fluorescence intensity of a GDI-1 reporter made from the related nematode C. briggsae (cdh3>GFP::Cbrgdi-1; n = 15 wild type and 13 gdi-1 RNAi treated animals; p = 0.586, Student's t-test; n.s = not significant). AU = arbitrary units; scale bar, 5 um.

Picture from Thirumaran P et al. (2024) MicroPubl Biol "Endogenous fluorescent reporters for heat shock proteins are not detectable ...."

Figure 1. Generation of endogenous reporters for heat shock proteins HSP-4 and HSP-60 and their validation with stress induction assays:(A) Schematic of CRISPR/Cas9 generated hsp-4::F2A::GFP::H2B and hsp-60::F2A::GFP::H2B tools. At the 3' end of hsp-4 and hsp-60 coding sequence, the following sequences were inserted using CRISPR/Cas9: a linker sequence (grey), F2A (yellow), GFP (green) and H2B (blue). (B) Heat shock increased the whole intestinal fluorescence (AU = arbitrary units) of both existing hsp-4p::GFP and hsp-60p::GFP transgenic reporters. For both tools, an approximate 4-fold increase in whole intestinal fluorescence was detected after heat shock. Representative images show an increase in fluorescence after heat shock. (C) GFP was not visibly induced in the intestinal nuclei of hsp-4::F2A::GFP::H2B and hsp-60::F2A::GFP::H2B tools. Representative images of both tools post-heat shock at 20x and 40x objectives show that GFP expression is not detected in intestinal nuclei (white circles). (D) Tunicamycin treatment significantly increased whole intestinal fluorescence (AU = arbitrary units) in the existing hsp-4p::GFP transgenic reporter. A significant increase was noted in intestinal fluorescence following tunicamycin treatment (n = 30). Representative images show the increase in fluorescence post-stress induction. (E) Paraquat treatment significantly increased whole intestinal fluorescence in the existing hsp-60p::GFP transgenic reporter. A significant increase was noted in intestinal fluorescence following paraquat treatment (n = 30). Representative images show the increase in fluorescence post-stress induction. (F) Endogenous hsp-4::F2A::GFP::H2B is not visibly induced in intestinal nuclei after tunicamycin treatment. Representative images of endogenous reporter hsp-4::F2A::GFP::H2B in GFP channel at 20x and 40x objectives show that no GFP expression is detected in intestinal nuclei (white circles). (G) Endogenous hsp-60::F2A::GFP::H2B is not visibly induced in intestinal nuclei after paraquat treatment. Representative images of endogenous reporter hsp-60::F2A::GFP::H2B in GFP channel at 20x and 40x objectives show that no GFP expression is detected in intestinal nuclei (white circles). P values were determined using unpaired Student's t-test. (H) Fluorescence of endogenous strains is not increased compared to wild-type in unstressed conditions. When unstressed, hsp-4::F2A::GFP::H2B and hsp-60::F2A::GFP::H2B whole intestinal fluorescence was not greater than wild-type (n = 30) suggesting that the fluorescence observed in the endogenous tools was due to background. P values were determined using one-way ANOVA. (I) The endogenous hsp-4::F2A::GFP::H2B and hsp-60::F2A::GFP::H2B reporters are not induced by heat stress. When exposed to heat stress (37oC, for 2 hours), hsp-4::F2A::GFP::H2B and hsp-60::F2A::GFP::H2B nuclear fluorescence is not induced. (n = 30). P values were determined using one-way ANOVA.

Picture from Nikonorova IA et al. (2022) Curr Biol "Isolation, profiling, and tracking of extracellular vesicle cargo in ...."

Figure 3. Nucleic acid binding transporter SID-2 is specifically enriched in the male- specific ciliated EV-releasing neurons (EVNs) and is shed in the form of EVs to the environment(A) Validation protocol for putative ciliary EV cargo. (B and C) EV release of SID-2::mScarlet and PKD- 2::GFP from cilia of CEM neuron of the head (B) and RnB neuron of the tail (C). (C) shows ray #7 in the process of shedding SID-2::mScarlet as evidenced by continuous fluorescence signal spanning the ciliary base, ciliary shaft, and the outside cloud of EVs. Other clouds on (C) are not labeled as EV release events because it was not clear from which cilium they originated. Resolution of the imaging technique was 140 nm in lateral di- rection and 400 nm in axial direction; thus, smaller EVs appear as clouds rather than individual EVs. Areas of intense inherent autofluorescence of the male tail are shaded with gray and labeled as au- toflu. Images show 3D renderings: on merged im- ages green and magenta colors represent different reporters, on images showing individual channels colors code for Z-depth-3.5 mm for (B) and 6.5 mm for (C).Ci, cilia; CEM, cephalic male neuron; HOB, hook neuron type B; RnB, ray neuron type B. Black and white maximum intensity projections of (B) and (C) and other data related to SID- 2::mScarlet expression are included in Figure S3.

Picture from Cevik S et al. (2013) PLoS Genet "Active transport and diffusion barriers restrict Joubert Syndrome-associated ...."

Figure 1. ARL-13/ARL13b localisation and mobility within an Inversin-like ciliary compartment. (A) Staining of mouse oviduct and tracheal tissue for endogenous Arl13b and acetylated tubulin shows proximal ciliary enrichment of Arl13b. Graph; line intensity profiles of Arl13b and acetylated tubulin (AcTub) signals from cilia denoted by white arrows. Bars; 5 mm (B) Co-expression of ARL-13::GFP with CHE-13/IFT57::mCherry, or ARL-13::tdTomato with either OSM-6::GFP or MKSR-1/B9D1::GFP, show that C. elegans ARL-13 is excluded from the transition zone (TZ). DS; distal segment. MS; middle segment. BB; basal body. PCMC; periciliary membrane compartment. Bars; 1 mm. (C) Staining of human hTERT-RPE1 cells shows that endogenous ARL13B does not colocalise with endogenous RPGRIP1L at the TZ. Bar; 10 mm (D) Phasmid cilia from L1 worms co-expressing ARL- 13::GFP with CHE-13/IFT57::mCherry show that the ARL-13 compartment extends to the ciliary tips in young larva. Graph shows ARL-13::GFP, KAP- 1::GFP (kinesin-II subunit) and OSM-6/IFT52::GFP ciliary compartment lengths in larval and adult stages of transgenic worms. Bar; 1 mm. (E) Fluorescence recovery after photobleaching (FRAP) curves after quenching 100%, or proximal-most 40%, of ARL-13::GFP ciliary signals in wild-type phasmid neurons. Signal ratio (au; arbitrary units) calculated from the average intensity of ARL-13 signal in the photobleached region compared to the non-photobleached region. All measurements are background subtracted and normalised to a pre-bleach ratio of 1.0. Each data point reports mean 6 SEM. (F) Time-lapse images taken from a recording of an amphid channel cilium from worms expressing ARL-13::GFP show processive retrograde movement of an ARL-13::GFP-associated particle. Kymograph and associated schematic derived from one such recording show multiple moving anterograde and retrograde particles. Bar; 1 mm.