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. 2019 Oct 21;29(20):3370–3385.e7. doi: 10.1016/j.cub.2019.08.017

Figure 2.

Figure 2

Gα12/13/Cta-GTP and Microtubules Control RhoGEF2 Enrichment at Cell Membrane in the Ectoderm

(A) Apical (0 μm), junctional (1.5 μm), and lateral (3.5 μm) z sections of ectoderm tissue co-expressing Myo-II::mCherry and GFP::RhoGEF2. Middle right panel, bottom left corner: a close up of a cell showing GFP::RhoGEF2 « comets » is shown (yellow arrowheads)

(B) Confocal z section of an ectodermal cell co-expressing RhoGEF2::RFP and EB1::GFP (1 z plan, 2 μm below the apical membrane). Both EB1 and RhoGEF2 « comets » co-localize; a slight spatial shift is observed between the two signals due to the acquisition condition (sequential acquisition).

(C) Apical (0 μm), junctional (1.5 μm), and lateral (3.5 μm) z sections of ectoderm tissue expressing GFP-RhoGEF2 in control, Gα12/13/Cta shRNA, and Gα12/13/CtaQ303L++ embryos. Note that the brightness of the GFP-RhoGEF2 signal has been artificially decreased in the Gα12/13/CtaQ303L++ panels for better visualization.

(D and E) Quantifications of mean medial-apical and junctional GFP::RhoGEF2 intensities in control, Gα12/13/Cta shRNA, and Gα12/13/Cta Q303L++ embryos. n, number of embryos. Medial and junctional GFP::RhoGEF2 intensities are decreased in Gα12/13/Cta shRNA embryos but are increased in Gα12/13/Cta Q303L++ embryos compared to control.

(F) Total GFP::RhoGEF2 cortical levels normalized to the apical junctional intensities (2 μm below the apical membrane).

(G) Confocal Z section of an ectodermal cell expressing GFP::RhoGEF2 in control and Gα12/13/CtaQ303L++ embryos (2 μm below the apical membrane). GFP::RhoGEF2 « comets » (yellow arrowheads) are absent from Gα12/13/CtaQ303L++ embryos.

(H) Confocal cross-section of an ectodermal cell (1 μm below apical membrane) expressing EB1::GFP in control and Gα12/13/CtaQ303L++ embryos.

Scale bars represent 5 μm. Means ± SEM between images are shown. Statistical significance has been calculated using Mann-Whitney U test. ns, p > 0.05; ∗p < 0.05; ∗∗p < 0.01. All the panels have the same orientation: dorsal at the top and anterior to the left.

See also Figures S2 and S7 and Videos S2 and S3.