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Figure 2


Fig. 2. Drosophila Cas is highly expressed in the embryonic nervous system. Localization of (A-D) DCas RNA and (E-H) protein reveals that DCas is highly expressed in the central and peripheral embryonic nervous system. (A,B) Stage 15 whole-mount Drosophila wild-type embryos viewed ventrally (A) and laterally (B) following in situ hybridization with antisense DCas RNA probes. Note the prominent DCas expression in the ventral nerve cord (VNC) and brain. Anterior, left. (C,D) A DCasP1/DCasP1 Drosophila embryo (C) subjected to in situ hybridization exhibits significantly reduced levels of DCas transcript, whereas an embryo deficient for the DCas gene (D) shows little to no DCas transcript (ventral views). (E) Western analysis of lysates obtained from late stage 16/17 embryos using an antibody generated against Cas (see text). The Cas antibody recognizes two prominent bands in wild-type embryo lysates (+) that are absent in DCas mutant lysates (Df). Molecular weight (100 kDa) is indicated. (F,G) A stage 15 wild-type embryo immunostained with Cas, showing Cas in axons within the anterior (A) and posterior (P) commissures, and longitudinal connectives (L) and motor nerves (asterisks). Cas immunostaining is also present at muscle attachment sites (arrows). (H) A stage 15 embryo homozygous for a DCas deficiency exhibits little to no Cas immunostaining (arrowheads, unstained longitudinal connective axons visualized using Nomarski optics). Scale bar: 30 µm in A-D; 10 µm in F-H.





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