Figure 4.
Apical-basal polarity regulates collagen IV–dependent cell–cell adhesion in the larval fat body. (A–D) Confocal single Z plane images of DAPI-stained, larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato +control (A), UAS-aPKC RNAi34332 (B), UAS-Crumbs RNAi39117 (C), and UAS-Scribble RNAi105412 (D; yellow or white arrowheads showing gaps at tricellular or bicellular cell–cell vertices, respectively). (E–L) Confocal images of larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato+Viking-GFP +control (E), +UAS-aPKC RNAi34332 (F), +UAS-Crumbs RNAi39117 (G), and +UAS-Scribble RNAi105412 (H; showing images of merged channels [single Z plane] and Viking-GFP channel [single Z plane and Z projection of 10 layers 2.5–5 μm from cell surface]; yellow or white arrow showing gaps at tricellular or bicellular vertices, respectively). Quantification of percentage of tricellular or bicellular cell–cell vertices containing gaps per image (I and J, respectively) from E–H (n: six images [control], nine images [UAS-aPKC RNAi34332], eight images [UAS-Crumbs RNAi39117], and eight images [UAS-Scribble RNAi10541], each from different animals). Kruskal–Wallis test followed by Dunn’s multiple comparisons, ****P < 0.0001, **P < 0.01, *P < 0.05, and ns P > 0.05. Quantification of mean CIVIC numbers (E′, F′, G′, and H′; from thresholded Z projection images of 10 Z planes of Viking-GFP channel [2.5–5 μm from cell surface], n: 10 tissues, 10 Z projection images per tissue and side, data paired by tissue) and mean intensity of Viking-GFP in the BM (E″, F″, G″, and H″; using surface ROIs in lateral view, data paired by tissue; n: six pupae [control], nine pupae [UAS-aPKC RNAi34332], eight pupae [UAS-Crumbs RNAi39117], and eight pupae [UAS-Scribble RNAi105412], 1 ROI per side for each tissue) on sides a and b. Paired two-sided t test, ****P < 0.0001. Quantification of CIVIC numbers (K) and mean intensity of Viking-GFP in the BM (L) for control, UAS-aPKC RNAi34332, UAS-Crumbs RNAi39117, and UAS-Scribble RNAi105412 on side b. Ordinary one-way multiple comparisons ANOVA, ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05, and ns P > 0.05. Scale bars; 20 µm (A–D and E–H).

Apical-basal polarity regulates collagen IV–dependent cell–cell adhesion in the larval fat body. (A–D) Confocal single Z plane images of DAPI-stained, larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato +control (A), UAS-aPKC RNAi34332 (B), UAS-Crumbs RNAi39117 (C), and UAS-Scribble RNAi105412 (D; yellow or white arrowheads showing gaps at tricellular or bicellular cell–cell vertices, respectively). (E–L) Confocal images of larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato+Viking-GFP +control (E), +UAS-aPKC RNAi34332 (F), +UAS-Crumbs RNAi39117 (G), and +UAS-Scribble RNAi105412 (H; showing images of merged channels [single Z plane] and Viking-GFP channel [single Z plane and Z projection of 10 layers 2.5–5 μm from cell surface]; yellow or white arrow showing gaps at tricellular or bicellular vertices, respectively). Quantification of percentage of tricellular or bicellular cell–cell vertices containing gaps per image (I and J, respectively) from E–H (n: six images [control], nine images [UAS-aPKC RNAi34332], eight images [UAS-Crumbs RNAi39117], and eight images [UAS-Scribble RNAi10541], each from different animals). Kruskal–Wallis test followed by Dunn’s multiple comparisons, ****P < 0.0001, **P < 0.01, *P < 0.05, and ns P > 0.05. Quantification of mean CIVIC numbers (E′, F′, G′, and H′; from thresholded Z projection images of 10 Z planes of Viking-GFP channel [2.5–5 μm from cell surface], n: 10 tissues, 10 Z projection images per tissue and side, data paired by tissue) and mean intensity of Viking-GFP in the BM (E″, F″, G″, and H″; using surface ROIs in lateral view, data paired by tissue; n: six pupae [control], nine pupae [UAS-aPKC RNAi34332], eight pupae [UAS-Crumbs RNAi39117], and eight pupae [UAS-Scribble RNAi105412], 1 ROI per side for each tissue) on sides a and b. Paired two-sided t test, ****P < 0.0001. Quantification of CIVIC numbers (K) and mean intensity of Viking-GFP in the BM (L) for control, UAS-aPKC RNAi34332, UAS-Crumbs RNAi39117, and UAS-Scribble RNAi105412 on side b. Ordinary one-way multiple comparisons ANOVA, ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05, and ns P > 0.05. Scale bars; 20 µm (A–D and E–H).

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