Figure 3.
Synthetic Rok kinase modulates mechanical properties of cells through the phosphorylation of Sqh::GFP and myosin II activity. (a) Schematic illustration of dorsal closure process in the developing fly embryo. Dorsal closure was used as a model to assess myosin II activation by means of Sqh::GFP phosphorylation with synthetic Rok. In sqh_Sqh::GFP embryos, expression of N-Rok::vhhGFP4 leads to abnormal dorsal closure. All panels show stills from live-imaging with dorsal views of the developing sqh_Sqh::GFP embryos at stages 13/14–16 (dorsal closure) expressing variants of the synthetic kinase in the engrailed domain (visualized by co-expression of mCherry-nls). Note the yellow arrows pointing to the Sqh::GFP foci and actomyosin cable invaginations in N-Rok::vhhGFP4 panel. (b) Stills from live imaging with dorsal views of the developing sqh_Sqh::GFP embryo expressing N-Rok::vhhGFP4 in the engrailed domain mounted with gluing to the coverslip technique to show a more severe dorsal open phenotype than embryos imaged on a glass-bottom dish shown in panel a. (c) Dorsal closure was used to compare N-Rok::vhhGFP4 with the previously published effectors that are known to activate myosin II. All panels show stills from live-imaging with dorsal views of the developing sqh_Sqh::GFP embryos at stages 13/14–16 (dorsal closure) expressing myosin II activating tool in the engrailed domain (visualized by co-expression of mCherry-nls). Only for Fig. 3, b and c and corresponding movies, the embryos were imaged with gluing to the coverslip technique; all the other presented embryos were imaged on a glass-bottom dish. Scale bars: 50 μm. Images are representative of n embryos indicated for every expressed synthetic kinase variant.

Synthetic Rok kinase modulates mechanical properties of cells through the phosphorylation of Sqh::GFP and myosin II activity. (a) Schematic illustration of dorsal closure process in the developing fly embryo. Dorsal closure was used as a model to assess myosin II activation by means of Sqh::GFP phosphorylation with synthetic Rok. In sqh_Sqh::GFP embryos, expression of N-Rok::vhhGFP4 leads to abnormal dorsal closure. All panels show stills from live-imaging with dorsal views of the developing sqh_Sqh::GFP embryos at stages 13/14–16 (dorsal closure) expressing variants of the synthetic kinase in the engrailed domain (visualized by co-expression of mCherry-nls). Note the yellow arrows pointing to the Sqh::GFP foci and actomyosin cable invaginations in N-Rok::vhhGFP4 panel. (b) Stills from live imaging with dorsal views of the developing sqh_Sqh::GFP embryo expressing N-Rok::vhhGFP4 in the engrailed domain mounted with gluing to the coverslip technique to show a more severe dorsal open phenotype than embryos imaged on a glass-bottom dish shown in panel a. (c) Dorsal closure was used to compare N-Rok::vhhGFP4 with the previously published effectors that are known to activate myosin II. All panels show stills from live-imaging with dorsal views of the developing sqh_Sqh::GFP embryos at stages 13/14–16 (dorsal closure) expressing myosin II activating tool in the engrailed domain (visualized by co-expression of mCherry-nls). Only for Fig. 3, b and c and corresponding movies, the embryos were imaged with gluing to the coverslip technique; all the other presented embryos were imaged on a glass-bottom dish. Scale bars: 50 μm. Images are representative of n embryos indicated for every expressed synthetic kinase variant.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal