Figure 7.
Neur-dependent disruption of the Crb complex is essential for NB ingression. (A) Loss of Sdt slows late ingression. Tukey’s box-and-whisker plots of NBs ingression speed (apical area reduction) in sdt-RNAi and controls embryos during early and late ingression. Medians (μm2/min) for control and sdt-RNAi: −1.3/−1.6 μm2/min (early ingression), ns, P = 0.44 and −0.6/−1.3 μm2/min (late ingression), ****, P = 1.9 × 10−5 (KS test). sdt-RNAi: 54 NBs, six embryos; controls: 30 NBs, four embryos. (B) Quantification of total apical fluorescence levels for all Sdt isoforms (Sdt::GFP, 36 NBs, six embryos) and for Sdt3 (Std3Sdt3::GFP, 48 NBs, six embryos). T = 0 min, onset of ingression; means ± SEM. (C) Ingressing live NBs in Sdt3::GFP (control) and SdtΔ3::GFP embryos (dots, arrowheads). Sdt3::GFP NBs lost their apical domain within ∼33 min (top panels, 48 NBs, four embryos). Apical plugs formed in SdtΔ3::GFP ingressing NBs (arrowheads in lower panels; 48 NBs, four embryos). Scale bar, 5 μm. (D) Mean apical area loss during NB ingression in Sdt3::GFP and SdtΔ3::GFP embryos. N values as in C. T = 0 min, onset of ingression; means ± SEM. (E) Quantification of apical area loss in three individual NBs from Sdt3::GFP (blue lines) and SdtΔ3::GFP (red lines) embryos during ingression. SdtΔ3::GFP NBs maintain an apical plug for 30–40 min after control cells have completed ingression. T = 0 min, onset of ingression. (F) Quantification of total Sdt3::GFP and SdtΔ3::GFP fluorescence at the apical domain during ingression. n values as in C. T = 0 min, onset of ingression; means ± SEM. (G) SdtΔ3::GFP embryos show reduced late ingression speed. Tukey’s box-and-whisker plots of ingression speed in Sdt3::GFP and SdtΔ3::GFP NBs during early and late ingression. Medians for Sdt3::GFP and SdtΔ3::GFP: −1.4/−1.3 μm2/min (early ingression), ns, P = 0.12 and −0.8469/−0.4946 μm2/min (late ingression), **, P = 1 × 10−3(KS test); n values as in C. (H) Co-localization of SdtΔ3::GFP and Crb at apical plugs in stage 9 SdtΔ3::GFP embryos (arrowheads). Scale bar, 5 μm. (I) Apical plugs in SdtΔ3::GFP embryos (arrowheads) do not contain Ecad. Dashed line indicates position of ZX cross-section through an apical plug shown in the right panel (I′) and schematic (I″). The apical plug containing SdtΔ3::GFP (arrowhead) is positioned apically to Ecad/AJs (arrow). Dlg labels basolateral membrane. Scale bars, 5 μm.

Neur-dependent disruption of the Crb complex is essential for NB ingression. (A) Loss of Sdt slows late ingression. Tukey’s box-and-whisker plots of NBs ingression speed (apical area reduction) in sdt-RNAi and controls embryos during early and late ingression. Medians (μm2/min) for control and sdt-RNAi: −1.3/−1.6 μm2/min (early ingression), ns, P = 0.44 and −0.6/−1.3 μm2/min (late ingression), ****, P = 1.9 × 10−5 (KS test). sdt-RNAi: 54 NBs, six embryos; controls: 30 NBs, four embryos. (B) Quantification of total apical fluorescence levels for all Sdt isoforms (Sdt::GFP, 36 NBs, six embryos) and for Sdt3 (Std3Sdt3::GFP, 48 NBs, six embryos). T = 0 min, onset of ingression; means ± SEM. (C) Ingressing live NBs in Sdt3::GFP (control) and SdtΔ3::GFP embryos (dots, arrowheads). Sdt3::GFP NBs lost their apical domain within ∼33 min (top panels, 48 NBs, four embryos). Apical plugs formed in SdtΔ3::GFP ingressing NBs (arrowheads in lower panels; 48 NBs, four embryos). Scale bar, 5 μm. (D) Mean apical area loss during NB ingression in Sdt3::GFP and SdtΔ3::GFP embryos. N values as in C. T = 0 min, onset of ingression; means ± SEM. (E) Quantification of apical area loss in three individual NBs from Sdt3::GFP (blue lines) and SdtΔ3::GFP (red lines) embryos during ingression. SdtΔ3::GFP NBs maintain an apical plug for 30–40 min after control cells have completed ingression. T = 0 min, onset of ingression. (F) Quantification of total Sdt3::GFP and SdtΔ3::GFP fluorescence at the apical domain during ingression. n values as in C. T = 0 min, onset of ingression; means ± SEM. (G) SdtΔ3::GFP embryos show reduced late ingression speed. Tukey’s box-and-whisker plots of ingression speed in Sdt3::GFP and SdtΔ3::GFP NBs during early and late ingression. Medians for Sdt3::GFP and SdtΔ3::GFP: −1.4/−1.3 μm2/min (early ingression), ns, P = 0.12 and −0.8469/−0.4946 μm2/min (late ingression), **, P = 1 × 10−3(KS test); n values as in C. (H) Co-localization of SdtΔ3::GFP and Crb at apical plugs in stage 9 SdtΔ3::GFP embryos (arrowheads). Scale bar, 5 μm. (I) Apical plugs in SdtΔ3::GFP embryos (arrowheads) do not contain Ecad. Dashed line indicates position of ZX cross-section through an apical plug shown in the right panel (I′) and schematic (I″). The apical plug containing SdtΔ3::GFP (arrowhead) is positioned apically to Ecad/AJs (arrow). Dlg labels basolateral membrane. Scale bars, 5 μm.

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