AKT1 phosphorylates Acn. (A–I) Micrographs of eyes expressing the indicated UAS transgenes under control of GMR-Gal4. Bar, 50 µm. (J) Western blots of anti-Myc immunoprecipitates of Myc-Acn^wt and Myc-Acn^S641,731A expressed in S2 cells and detected with antibodies against Acn or Acn-pS641. S2 cell lysates were probed for Actin. IP, immunoprecipitation; wt, wild type. (K) Western blots of lysates from S2 cells expressing Myc-Acn^wt that were treated without (−) or with calf intestinal phosphatase (CIP) and probed for Acn, Acn-pS641, or Actin. (L–M″) Apical most regions of eye discs that include cone cell nuclei (marked CC) are depicted in z projections of confocal sections. Eye discs were from larvae expressing Myc-Acn^wt (L–L″) or Myc-Acn^S641,731A (M–M″) and were stained for Acn-pS641, Myc, and DNA. Arrows point to unspecific staining in mitotic cells. (N–N″) Projection of confocal sections of eye disc expressing AKT1-RNAi within RFP-marked clones (magenta) and stained for Acn-pS641 (N′) and DNA (N″). The broken line indicates the clone boundary. Detailed genotypes and roughness quantification are given in Tables S1 and S2.