Fig. 5. The Skd and Kto proteins associate in vivo. (A-F) Third instar eye discs. (A-C) show skd^T413-mutant clones. Wild-type tissue is labeled by anti-ß-gal staining reflecting arm-lacZ expression (green; B,C). Rabbit anti-Skd staining (red; A,C). (D-F) kto^T241-mutant clones. Wild-type tissue is labeled with GFP (green; E,F). Rat anti-Kto staining (red; D,F). In both cases, the antibodies do not stain mutant tissue, but show ubiquitous nuclear staining in wild-type tissue. The same pattern was observed for the guinea pig anti-Kto antibody (not shown). (G-H) Coimmunoprecipitation of Skd, Kto and SOH1. (G) Coimmunoprecipitation of Kto and Soh1 with an anti-Skd antibody. The upper gel is blotted with rat anti-Kto and the lower gel with anti-SOH1. Lane 1, 10% white^- total embryonic extract; lane 2, IP with anti-Skd from white^- extract; lane 3, control IP from white^- extract; lane 4, IP with anti-Skd from extract overexpressing UAS-skd and UAS-kto with da-GAL4; lane 5, control IP from extract overexpressing UAS-skd and UAS-kto with da-GAL4. (H) Coimmunoprecipitation of Skd and Soh1 with the guinea pig anti-Kto antibody. The upper gel is blotted with anti-Skd and the lower gel with anti-SOH1. Lane 1, 2.5% white^- total extract; lane 2, IP with anti-Kto from white^- extract; lane 3, control IP from white^- extract; lane 4, IP with anti-Kto from extract overexpressing UAS-skd and UAS-kto with da-GAL4; lane 5, control IP from extract overexpressing UAS-skd and UAS-kto with da-GAL4.

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