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[Preprint]. 2025 Jan 28:2023.11.20.567915. Originally published 2023 Nov 20. [Version 3] doi: 10.1101/2023.11.20.567915

Fig. 2. UNC-61a TMD is required for normal vulval morphology and function.

Fig. 2.

(A) (top) Schematic of adult C. elegans hermaphrodite (pink: pharynx, blue: germline, orange: vulva.) (bottom) Images are inverted contrast of the fluorophore annotated. Arrow: vulval epithelium apical surfaces; arrowhead; linear septin decorated structures. Dashed lines outline the C. elegans terminal bulb and oogenic germline. The fluorescent signal beneath the germline corresponds to the autofluorescence of the C. elegans intestine. Scale bar = 10 μm (pharynx and vulva); 25 μm (germline). (B) (top) Schematic of representative images of protruded vulva (pvl) phenotypes and embryos in uterus. (middle) Representative transmitted light images of pvl phenotypes. Arrow denotes embryos that are more developed than they should be indicating an egg-laying defective (egl) phenotype. (bottom) Percent incidence of post-embryonic phenotypes (pvl: protruded vulva; egl: egg-laying defective; bag: bag-of-worms) of control (grey), unc-61Δ (pink), unc-61 Δa (yellow), and unc-61a ΔTMD (blue) animals (n=100 animals). (C) (left top) Schematic of control and unc-61a ΔTMD vulvae (green: epithelium, black: apical surface, grey: surrounding muscles.) (left bottom) Inverted contrast images of Lifeact::GFP (F-actin) in vulva region. Arrowhead: muscle cuticle connection in control, disconnection in unc-61a ΔTMD; arrows: apical surface of the vulval epithelium. (right) Percent incidence of muscle detachment and open vulva phenotypes. “n” above bar indicates the number of animals scored for each condition. Scale bar = 20 μm. (D) (left) Representative images of uterine and embryo permeability in control and unc-61a ΔTMD animals. Arrowheads denote vulval location, asterisks indicate germline location, and dashed lines outline uterus. Scale bar = 50 μm. (right) Quantification of uterine (dashed lines) mean fluorescence intensity in control (grey; n = 15 animals) and unc-61a ΔTMD animals (blue; n = 15 animals). Error bars are mean ± SD. ****, ≤ 0.0001, as determined by Student’s unpaired t-test.