Picture from Lai CC et al. (1996) J Cell Biol "Sequence and transmembrane topology of MEC-4, an ion channel subunit required ...."

Figure4. MEC-4 is expressed in the six touch receptor neurons and precluded from the nucleus. (A) Immunoreactivity in a whole mount preparation of strain ZB2 incubated with anti-NH2-terminal MEC-4 antibody AbM4(1-69). ZB2 harbors an integrated ar- ray of a mec-4 lacZ fusion gene (Mitani et al., 1993). Indicated are the six touch receptor neurons. This photograph is a composite of three photographs taken to view different touch cells in the same focal plane. (B) Detail of anti-MEC-4 immunoreactivity in the two tail touch receptors, PLML and PLMR. (C) DAPI-stained nuclei in the tail of the animal depicted in B. (Arrows) Touch receptor nuclei. Overlay of photographs in B and C indicates that the dark inner area typically seen in stained cells corresponds to the nucleus.

Picture from Durkan, Aidan et al. (2022) MicroPubl Biol

Figure 1. RNAi knockdown of dcaf-13 impairs C elegans growth, fertility, and development.:A-C) Representative images of 2-day old A) L4440 control B) Ahringer library dcaf-13(RNAi) or C) Vidal library dcaf-13(RNAi) treated C. elegans. Images were taken at 25X magnification, scale bar=0.25mm. D) Quantification of worm lengths for 2-day old L4440 control or dcaf-13 RNAi treated C. elegans. Graph shows combined data from 3 independent biological replicates, n=10 for each replicate. Treatment with both Ahringer (AR) and Vidal library dcaf-13 RNAi clones resulted in a significant reduction in worm length compared to L4440 control (One-way ANOVA with Sidak's multiple comparisons, p<0.001). E) Total progeny produced by 3 adult worms during a 12-hour laying period. L4 stage worms were treated with control or dcaf-13 RNAi for 24 hours, then allowed to lay eggs for 12 hours before being removed. 24 hours after adult removal, embryos and larvae were counted. Treatment with both AR and Vidal dcaf-13 RNAi resulted in less progeny production compared to L4440 control treatment (One-way ANOVA with Sidak's multiple comparisons, n=4, p<0.05). Data shown is from four independent biological replicates. F) Percent embryonic lethality was calculated by dividing the number of unhatched embryos by the total number of unhatched embryos and live larvae produced by RNAi or control treated worms at least 24 hours after egg laying and adult removal. No statistically significant differences in percent embryonic lethality were found (Kruskal-Wallis test, n=4, p=0.08). Data shown is from four independent biological replicates. G) Larvae produced during a 12-hour egg laying window by dcaf-13 RNAi or control treated C. elegans were followed for 72 hours after egg laying and the number of larvae to have reached L4 stage at each time point was recorded. None of the dcaf-13(RNAi) larvae had reached L4 stage as of 72 hours post egg laying. Data shown is the average of 2 independent biological replicates. *p<0.05, ****p<0.001

Picture from Hoffmann M et al. (2009) PLoS One "C. elegans ATAD-3 is essential for mitochondrial activity and ...."

Figure 2. ATAD-3 is a mitochondrial protein and its depletion leads to arrest at developmental stages with low mitochondrial activity. (A) Western Blot analysis of C. elegans wild type cytoplasmic and mitochondrial protein fraction using anti-NUO-2, anti-ATAD-3 and anti-GAPDH antibodies to identify mitochondrial NUO-2/ATAD-3 (NUO-2 = homologue of the human NADH ubiquinone oxidoreductase subunit NDUFS3) and cytoplasmic GAPDH (glyceraldehyde-3-phosphate dehydrogenase), respectively. (B) Western blot analysis of protein extracts from wild type (WT) and atad-3(RNAi) animals reveals a strongly reduced ATAD-3 signal. For comparison of relative protein levels, GAPDH served as a loading control. (C,D) Mitochondrial morphology was analyzed in transgenic strain SJ4103 expressing a mitochondrial targeted GFP under control of the muscle-specific myo-3 promoter ([25]). Bar: 10 um; N = nucleus. (C) In SJ4103 control animals (fed with HT115 bacteria carrying the empty L4440 feeding-vector), mitochondria in body wall muscle form a well organized interconnected reticulum. (D) After atad-3(RNAi), mitochondria appear disorganized and thinner. (E) Quantification of mitochondrial filamentation (aspect ratio = AR) and mitochondrial branching (formfactor = F) revealed no significant changes. Number of individual animals analyzed: WT = 11, atad-3(RNAi) = 11; number of objects analyzed: WT = 780, atad-3(RNAi) = 797. (F) Enzymatic activities (nmol/min/mg) of complex I (CI) and citrate synthase (CS) of a mixed stage WT population (green), WT L1 larvae (yellow) and atad-3(RNAi) animals (red). Asterisks indicate significant differences (p<0.05) in comparison to the control.

Picture from Shostak Y et al. (2004) Genes Dev "Identification of C. elegans DAF-12-binding sites, response elements, and ...."

Figure 4. DAF-12 regulates LIT-1 gene expression in C. el- egans. In A-C, the wild-type or m11m31 mutant 4.2 fragment was subcloned upstream of a minimal pes-10 promoter driving GFP expression and microinjected into wild-type or daf-12 mu- tant rh61rh412 C. elegans. (A) GFP expression in the pharynx (arrows) in N2 worms with wild-type 4.2. (B) Absence of GFP pharyngeal expression in N2 worms injected with the 4.2m11m31 mutant reporter. One-hundred percent of animals exhibited the effect. (C) Reduced GFP expression in daf-12 mu- tant worms (arrows) injected with wild-type 4.2 fragment. In four independent injections, >90% of animals exhibited the ef- fect in three cases; in the fourth, 50% displayed the reduction. (D-L) A 4-kb LIT-1 genomic segment (nucleotides 4527-8561 from cosmid W06F12, with upstream/intronic sequences in red and exons in blue) with the LIT-1b initiating methionine codon (M) at its midpoint, and bearing either the wild-type or m11m31 4.2 segment in its second intron (nucleotides 7201-7408), was fused in frame to GFP and microinjected into wild-type or daf- 12 mutant rh61rh412 C. elegans. (D-F) LIT-1-GFP expression in pharynx (arrows) with wild-type or m11m31 4.2 constructs in L4 N2 worms, or with wild-type 4.2 constructs in daf-12 mu- tant worms, respectively. In E, 100% of animals had no pharyn- geal GFP expression; in F, >90% of animals had reduced GFP expression in pharynx. (G) Low levels of LIT-1-GFP expression from 4.2 construct in seam cells of L4 N2 animals (arrow); GFP expression in embryos served as positive control. (H,I) Elevated LIT-1-GFP expression in seam cells (arrows) with m11m31 4.2 constructs in L4 N2 worms, or with wild-type 4.2 constructs in daf-12 mutant worms, respectively. Five times more animals than in G had elevated expression. (J) Low levels of LIT-1-GFP expression from 4.2 construct in vulva cells (arrow) of L4 N2 animal. (K,L) Elevated LIT-1-GFP expression in vulva cells (ar- rows) with m11m31 4.2 constructs in L4 N2 worms, or with wild-type 4.2 constructs in daf-12 mutant worms, respectively. Five times more animals than in J had elevated expression. In A-L, >50 animals per panel were examined. In A-F and J-L, images were taken at 1000×; in G-I, images were taken at 400×. (M) Regulation of endogenous LIT-1 expression by DAF-12 in L4. LIT-1a and LIT-1b mRNAs were analyzed by qRT PCR; data were normalized to AMA-1 abundance values, which were as- signed a value of 100. CAP-1, CAP-2, and RGR-1 were used as controls; various other control gene transcripts, including ACT- 5, RPL-19, and DPY-26, also did not change (data not shown).