Destabilisation of E-APC in sgg mutant embryos. (a, b) Face-on and (c) side views of ~14 hours old embryos (stage 16), fixed and co-stained with antibodies against GFP and α-catenin as indicated, revealing junctional association of GFP-E-APC in (a) wild-type and (b, c) sgg mutant embryos (similar in sgg null and paternally rescued embryos, see also Fig. 4f). (d, e) Western blots of hand-picked 10–16 hours old wild-type and sgg mutant embryos (~100 embryos per lane), probed with antibodies against (d) GFP or (e) E-APC, and α-tubulin as internal controls. Note that the levels of GFP-E-APC and of endogenous E-APC are much reduced in sgg compared to wild-type embryos (sgg mutants represent a 1:1 mixture of sgg null and paternally rescued embryos). The lower bands in upper panels (d, e) correspond to breakdown products of GFP-E-APC and E-APC, respectively; their occurrence varies somewhat between preparations.