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Fig. 6. Cos2-572A and Cos2-572D do not rescue cos2-null phenotypes in embryos. (A-C) Wild-type embryos. (D-O) cos2^w1 germline clones (glc; D-F) expressing UAS-cos2-WT (G-I), UAS-cos2-572A (J-L) or UAS-cos2-572D (M-O) under the control of the ubiquitous 69BGal4 driver. (A,D,G,J,M) Cuticle views of first instar larva. (Other panels) In blue, mRNA in situ hybridisation for wg (B,E,H,K,N) and rho (C,F,I,L,O). In brown, immunostaining against En. The wild-type embryo displays an alternating segment pattern of naked cuticle and denticle belts composed of six types of denticles numbered from 1 to 6 (A). In cos2-null embryos, an anterior expansion of wg (E) and a posterior expansion of rho (F) are observed, and only naked cuticle and denticle types 1 and 2 are made (D). Expression of a cos2-WT transgene fully rescues the cos2^w1 glc phenotype defects with restored cuticle pattern and correct Hh target gene expression (G-I). By contrast, the rescue is much weaker with the cos2-572A transgene, and is barely observable with cos2-572D. Embryos were raised at similar temperatures (18°C). Two to three different transgenic lines were tested for each variant in rescue experiments and showed comparable results. (P) Western blot for Cos2-Myc and Arm proteins from embryos expressing cos2-WT-myc, cos2-572A-myc or cos2-572D-myc driven by the armGal4 driver. (Q) Bar graph showing the quantity of Cos2-Myc as analysed by western blot as in P, expressed as a percentage relative to the amount of Arm. The mean of two independent experiments is shown.

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