Figure 8.

Co-immunoprecipitation of PPE proteins. A shows an experiment where HA-In and Fy-Flag were co-immunoprecipitated by either anti-In or anti-Flag antibodies. In this experiment a heat shock was used to induce the expression of transgenes subcloned behind the hsp70 promoter. B shows the co-immunoprecipitation of In and Frtz. This experiment used UAS-HA-in and UAS-myc-frtz transgenes driven by ptc-Gal4. Wing disc samples were immunoprecipitated using Rabbit anti-HA antibody. Arrows point to myc-Frtz and HA-In on the Western blots. C shows an experiment where we tested the co-immunoprecipitation of Fy and Frtz. Samples from ptc-Gal4/UAS-fy-Flag-Ollas; UAS-myc-frtz/+ wing discs were IP with anti-Myc antibody and then assayed by western blotting. Lane 3 and 4 (on the left side) are duplicates. The arrows point to Fy-Flag-Ollas (left) and myc-Frtz (right). Note that Fy was not preciptiated with Frtz. D shows an experiment where all three of the PPE proteins were co-expressed (ptc-Gal4/UAS-HA-in UAS-fy-GFP; UAS-frtz-GFP/+). Samples were made from wing discs and immunoprecipitated using anti-Fy antibody. The Western blots were probed with anti-GFP (to detect both Fy-GFP and Frtz-GFP) and anti-In. Control experiments established that anti-Fy antibody could not pull down either HA-in or Frtz-GFP (data not shown).