Figure 3.
Excess apical aPKC and apical constriction in Cactin knockdown cells. (A–D‴) Images of anti-Arm and anti-aPKC co-staining of posterior follicle cells with c306-Gal4 driving UAS-LifeAct-GFP together with (A–B‴) control (crossed to w1118) or (C–D‴) UAS-cactin-RNAi. (A, A′, C, and C′) Single section view. (B–B‴ and D–D‴) Projection view. Note that in B‴ and D‴, to show the DNA signal in follicle cells more clearly, the autofluorescence from the yolk was masked in black. (E–F′ and G–H′)(E–F′) Anti-aPKC staining and (G–H′) anti-Arm of HS-flp-out clones showing the apical constriction phenotypes in Cactin knockdown follicle cells. (E, E′, G, and G′) Control (UAS-w-RNAi). (F, F′, H, and H′) UAS-cactin-RNAi. Arrowhead indicates the apical domain in a cactin-RNAi clone. (E′ and F′) Images show the aPKC channel in E and F. (G′ and H′) Images show the Arm channel in G and H. (I) Quantification of aPKC staining intensity in HS-flp-out clones showing the apical constriction phenotypes. The ratio of aPKC staining intensity in RNAi clones (GFP+ cells)/non-RNAi clones (GFP− cells) is shown. Ratio = 1 suggests that there is no difference between the RNAi clones and the non-RNAi clones. Ratio > 1 suggests RNAi clones have higher levels of aPKC than non-RNAi clones. (J–K‴) HS-flp-out clones showing anti-aPKC and anti-mCherry co-staining in follicle cells. GFP (green) labels Gal4-expressing clones. (J–J‴) Control (UAS-w-RNAi). (K–K‴) UAS-cactin-RNAi. Arrowhead indicates the aPKC is concentrated in Cactin knockdown cells. (J′–J‴ and K′–K‴) Images show the single channels in J and K. (L) Quantification of aPKC staining intensity in HS-flp-out clones. The ratio of aPKC staining intensity in RNAi clones (GFP+ cells)/non-RNAi clones (GFP− cells) is shown. (M–P) Images of anti-aPKC and anti-Dlg co-staining border cell clusters with c306-Gal4 together with (M) control (cross to w1118) or (O) UAS-cactin-RNAi. (M′, M″, O′, and O″) Images show the single channels in M and O. (N–P) Schematic of polarity of the border cell cluster in M–O. (Q–R′) Images of anti-aPKC and anti-Crb co-staining border cell clusters with c306-Gal4 together with (Q) control (cross to w1118) or (R) UAS-cactin-RNAi. (Q′ and R′) Images show the anti-Crb single channels in Q and R. Yellow arrowheads indicate the apical junctions in border cell clusters. Scale bars, 20 μm.

Excess apical aPKC and apical constriction in Cactin knockdown cells. (A–D‴) Images of anti-Arm and anti-aPKC co-staining of posterior follicle cells with c306-Gal4 driving UAS-LifeAct-GFP together with (A–B‴) control (crossed to w1118) or (C–D‴) UAS-cactin-RNAi. (A, A′, C, and C′) Single section view. (B–B‴ and D–D‴) Projection view. Note that in B‴ and D‴, to show the DNA signal in follicle cells more clearly, the autofluorescence from the yolk was masked in black. (E–F′ and G–H′)(E–F′) Anti-aPKC staining and (G–H′) anti-Arm of HS-flp-out clones showing the apical constriction phenotypes in Cactin knockdown follicle cells. (E, E′, G, and G′) Control (UAS-w-RNAi). (F, F′, H, and H′) UAS-cactin-RNAi. Arrowhead indicates the apical domain in a cactin-RNAi clone. (E′ and F′) Images show the aPKC channel in E and F. (G′ and H′) Images show the Arm channel in G and H. (I) Quantification of aPKC staining intensity in HS-flp-out clones showing the apical constriction phenotypes. The ratio of aPKC staining intensity in RNAi clones (GFP+ cells)/non-RNAi clones (GFP− cells) is shown. Ratio = 1 suggests that there is no difference between the RNAi clones and the non-RNAi clones. Ratio > 1 suggests RNAi clones have higher levels of aPKC than non-RNAi clones. (J–K‴) HS-flp-out clones showing anti-aPKC and anti-mCherry co-staining in follicle cells. GFP (green) labels Gal4-expressing clones. (J–J‴) Control (UAS-w-RNAi). (K–K‴) UAS-cactin-RNAi. Arrowhead indicates the aPKC is concentrated in Cactin knockdown cells. (J′–J‴ and K′–K‴) Images show the single channels in J and K. (L) Quantification of aPKC staining intensity in HS-flp-out clones. The ratio of aPKC staining intensity in RNAi clones (GFP+ cells)/non-RNAi clones (GFP− cells) is shown. (M–P) Images of anti-aPKC and anti-Dlg co-staining border cell clusters with c306-Gal4 together with (M) control (cross to w1118) or (O) UAS-cactin-RNAi. (M′, M″, O′, and O″) Images show the single channels in M and O. (N–P) Schematic of polarity of the border cell cluster in M–O. (Q–R′) Images of anti-aPKC and anti-Crb co-staining border cell clusters with c306-Gal4 together with (Q) control (cross to w1118) or (R) UAS-cactin-RNAi. (Q′ and R′) Images show the anti-Crb single channels in Q and R. Yellow arrowheads indicate the apical junctions in border cell clusters. Scale bars, 20 μm.

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