Figure 10. Chitin levels modulate Src activation.

(A and D) Wild-type embryo expressing AbdBGAL4UASkkvRNAi and UASSrcGFP, stained with anti-GFP (green, A) to visualise the AbdB domain, anti-pSrc (red, A’) to detect activated Src42A, and fluostain (grey, A’’) to label chitin. Levels of pSrc are better visualised in D, with a different colour-coded LUT. (B and E) Blimp-1 mutant embryo, expressing AbdBGAL4UASBlimp-1 and UASSrcGFP, to rescue the Blimp-1 phenotype at the posterior end of the embryo, stained with anti-GFP (green, B) to visualise the AbdB domain, anti-pSrc (red, B’) to detect activated Src42A, and fluostain (grey, B’’) to label chitin. Levels of pSrc are better appreciated in E, with a different colour-coded LUT. (C and F) kkv mutant embryo, expressing AbdB-GAL4 and UASkkvGFP, to rescue the kkv phenotype at the posterior end of the embryo, stained with anti-GFP (green, C) to visualise the AbdB domain, anti-pSrc (red, C’) to detect activated Src42A and fluostain (grey, C’’) to label chitin. Levels of pSrc are better appreciated in F, with a different colour-coded LUT. Scale bars 10 µm. (G-I) Late stage 16 embryos stained with fluostain to visualise taenidial organisation. (G) Detail of Blimp-1 dorsal trunk showing taenidial disorganisation; (H) whole Src42A;Blimp-1 double mutant, showing some of the characteristic Blimp-1 tracheal phenotypes; (I) Detail of Src42A;Blimp-1 double mutant dorsal trunk showing rescued taenidial organisation. Scale bars 25 µm.

DOI: http://dx.doi.org/10.7554/eLife.09373.019

Figure 10—figure supplement 1. Src protein levels in kkv mutant trachea.

(A, B) Wild-type embryo expressing AbdBGAL4UASkkvRNAi stained with anti-Src (red, A and A’, B and B’) to detect Src42A protein and anti-DE-Cad (green, A and A’’, B and B’’). In panel A we also detected chitin with fluostain. (A) is a projection of confocal sections and (B) is a single section. (C) kkv heterozygous embryo stained with anti-Src (red, C and C’) to detect Src42A protein and anti-DE-Cad (green, C and C’’). Image is a single confocal section. (D) kkv homozygous embryos stained with anti-Src (red, D and D’) to detect Src42A protein and anti-DE-Cad (green, D and D’’). Image is a single confocal section. (E) Src42A heterozygous embryo stained with anti-pSrc (red, E and E’) to detect phosphorylated Src42A protein and CBP (blue, E). Image in E is a projection of confocal sections and in E’ is a single confocal section. (F) Src42A homozygous embryo stained with anti-pSrc (red, F and F’) to detect phosphorylated Src42A protein and CBP (blue, F). Images in F and F’ are projections of confocal sections.

DOI: http://dx.doi.org/10.7554/eLife.09373.020