Figure 5.
Tracheal system of Drosophila embryo develops abnormally due to the excessive myosin II phosphorylation by engineered N-Rok kinase. (a) All panels show lateral views of fixed sqh_Sqh::GFP embryos at stages 14–16 expressing the indicated variant of synthetic Rok kinase in the tracheal system (btl expression domain visualized by co-expression of mCherry-nls). Embryos were stained with an anti-phospho-Sqh/MRLC antibody. For every expressed synthetic kinase, the number of embryos analyzed is indicated (n). For N-Rok::dGBP1, the n number is given as a proportion of embryos in which clear p-Sqh clumps corresponding to Sqh::GFP foci were visible, to the total number of embryos included in the analysis. Scale bar, 20 μm. (b) Aberrant tracheal development in sqh_Sqh::GFP embryos expressing the indicated variant of synthetic Rok kinase in the tracheal system. All panels show stills from live imaging with lateral views of sqh_Sqh::GFP embryos at stages 11/12, 14, and 17, corresponding to the stages depicted on the schematic drawing above. The panel on the right shows magnification of the indicated region from the last stage. Yellow arrows point to Sqh::GFP foci visible already at stage 11/12; asterisks indicate clusters of tracheal cells around the Sqh::GFP foci. Scale bars, 50 μm; in the zoomed panel, 20 μm.