Figure 3.
E-cadherin RNAi is not sufficient to cause cell–cell adhesion defects in the larval fat body. (A–B‴) Confocal images of CAAX-GFP–expressing larval fat body immunostained for Baz (A and A′), and E-cadherin (B, B′; side a [top] and b [bottom] in planar view [A and B] and lateral view [A′ and B′], blue and orange arrowheads pointing at cell surfaces or lateral domains, respectively). Quantification of mean intensities of Baz (A″ and A‴) and E-cadherin (B′ and B‴) on surfaces (‘, yellow) or lateral domains (‘’, orange) on side a and b (mean of mean intensities from several ROIs, data paired by tissue; n: 10 tissues, 10 or 5 surface or lateral ROIs per side for Baz or E–Cadherin, respectively [A″–B″ and A‴–B‴]). Paired two-sided t test, ****P < 0.0001. (C–E) Confocal images of larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato +control (C) or +UAS-E-cadherin RNAi32904 (D) immunostained for E-cadherin (side a and b shown in planar [top] and lateral views [bottom], black arrowhead pointing at cell–cell vertex, blue and orange arrowheads pointing at surface or lateral domain, respectively). Note that the same control images are displayed in Fig. 3 C and Fig. S2 A. Fig. 3, C–E and Fig. S2, A–C′ are the results from the same experiment and hence the control is the same for both. Quantification of mean intensity of E-cadherin for control (C′) or UAS-E-cadherin RNAi32904 (D′) on surfaces on sides a and b (mean of mean intensities from several ROIs, data paired by tissue; n: three tissues, three surface ROIs per side). Unpaired two-sided t test, ****P < 0.0001. Quantification of mean intensities of E-cadherin for control, UAS-E-cadherin RNAi32904 (from C′ and D′), UAS-E-cadherin RNAi103962, and UAS-E-cadherin RNAi27082 (from Fig. S2, B′ and C′) shown for side a (E). Ordinary one-way multiple comparisons ANOVA, ****P < 0.0001. (F and F′) Confocal images of larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato+Viking-GFP (F; side a and b shown in planar [top] and lateral views [bottom], black and white arrowheads pointing at CIVICs at cell–cell vertices; merge and single channels of Z projection of 10 Z planes at 2.5–5 μm from cell surface). Quantification of CIVIC numbers per image (F′, using 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). Paired two-sided t test, ****P < 0.0001, ns P > 0.05. Scale bars: 20 µm (A–D and F).

E-cadherin RNAi is not sufficient to cause cell–cell adhesion defects in the larval fat body. (A–B‴) Confocal images of CAAX-GFP–expressing larval fat body immunostained for Baz (A and A′), and E-cadherin (B, B′; side a [top] and b [bottom] in planar view [A and B] and lateral view [A′ and B′], blue and orange arrowheads pointing at cell surfaces or lateral domains, respectively). Quantification of mean intensities of Baz (A″ and A‴) and E-cadherin (B′ and B‴) on surfaces (‘, yellow) or lateral domains (‘’, orange) on side a and b (mean of mean intensities from several ROIs, data paired by tissue; n: 10 tissues, 10 or 5 surface or lateral ROIs per side for Baz or E–Cadherin, respectively [A″–B″ and A‴–B‴]). Paired two-sided t test, ****P < 0.0001. (C–E) Confocal images of larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato +control (C) or +UAS-E-cadherin RNAi32904 (D) immunostained for E-cadherin (side a and b shown in planar [top] and lateral views [bottom], black arrowhead pointing at cell–cell vertex, blue and orange arrowheads pointing at surface or lateral domain, respectively). Note that the same control images are displayed in Fig. 3 C and Fig. S2 A. Fig. 3, C–E and Fig. S2, A–C′ are the results from the same experiment and hence the control is the same for both. Quantification of mean intensity of E-cadherin for control (C′) or UAS-E-cadherin RNAi32904 (D′) on surfaces on sides a and b (mean of mean intensities from several ROIs, data paired by tissue; n: three tissues, three surface ROIs per side). Unpaired two-sided t test, ****P < 0.0001. Quantification of mean intensities of E-cadherin for control, UAS-E-cadherin RNAi32904 (from C′ and D′), UAS-E-cadherin RNAi103962, and UAS-E-cadherin RNAi27082 (from Fig. S2, B′ and C′) shown for side a (E). Ordinary one-way multiple comparisons ANOVA, ****P < 0.0001. (F and F′) Confocal images of larval fat body expressing Lpp-Gal4+UAS-Myr-td-Tomato+Viking-GFP (F; side a and b shown in planar [top] and lateral views [bottom], black and white arrowheads pointing at CIVICs at cell–cell vertices; merge and single channels of Z projection of 10 Z planes at 2.5–5 μm from cell surface). Quantification of CIVIC numbers per image (F′, using 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). Paired two-sided t test, ****P < 0.0001, ns P > 0.05. Scale bars: 20 µm (A–D and F).

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