Cyst physically interacts with Baz and the Crb complex. (A) Z-projections taken from live stage-9 embryo expressing GFP::Cyst^R. Embryos have been injected with dsRNAs against crb or baz (n = 40 each). GFP::Cyst^R is lost from the cortex in knockdown embryos. Scale bar, 10 µm. (B) Immunoprecipitation (IP) with anti-GFP followed by immunoblot (IB) with anti-FLAG. Cyst constructs were coexpressed with Baz::FLAG and Patj::FLAG in HEK293T cells. Input shows Baz::FLAG and Patj::FLAG bands. GFP::Cyst^R and GFP::Cyst^RΔPBM formed Baz::FLAG- or Patj::FLAG-containing complexes. GFP::Cyst^R-C and Patj::FLAG formed a particularly strong complex. GFP::Cyst^R-C and Baz::FLAG formed a weak complex. (C) Immunoprecipitation with anti-GFP followed by immunoblot with anti-FLAG. A FLAG-tagged version of Cyst C-terminal region (FLAG::Cyst^R-C) was coexpressed with various GFP-tagged fragments of Cyst (or GFP as a control) in HEK293T cells. Input shows FLAG::Cyst^R-C. GFP::Cyst^R, GFP::Cyst^RΔPBM, and GFP::Cyst^R-C formed FLAG:: Cyst^R-C–containing complexes. (D) Controls for B and C. Immunoprecipitation with anti-GFP followed by immunoblot with anti-GFP. All GFP-tagged constructs are expressed.