Strip or Cka depletion has little effect on total moesin and E-cadherin (shotgun) levels and distribution. (A) X–z section along the anterior– posterior axis of a wing disc expressing P35 together with dsRNA targeting Cka in a central stripe of cells under the control of ptc-GAL4. The disc was stained with DAPI (blue) to visualize nuclei and with anti–p-moesin (green) and anti-Ptc (blue) antibodies. The image was generated by orthogonal projection. Apical p-moesin levels are decreased specifically in RNAi-expressing cells in the ptc domain. (B–G) Confocal projections through medial portions of the hinge region of wing discs expressing GFP (green) in a stripe of cells under control of ptc-GAL4, either alone (B and E) or together with dsRNA targeting Strip (C and F) or Cka (D and G). Discs were stained with DAPI (blue) to visualize nuclei and with antibodies against moesin (red) and Shg (shown separately in E–G). Yellow dotted lines indicate limits of the GFP-transgene expression domains. (H–M) Quantification of normalized total moesin (H–J) and Shg (K–M) fluorescence intensity in hinge cells expressing GFP alone (H and K) or together with dsRNA targeting Strip (I and L) or Cka (J and M) compared with their respective adjacent wild-type (GFP−) cells. Fluorescence was measured at multiple folds in five wing discs for each genotype. Two-tailed paired sample t test with the following P values: NS, P > 0.05; *, P < 0.05. RNAi expression had little effect on moesin and Shg levels and distribution, with only a small increase (4%) in moesin levels in Cka-depleted cells and a small decrease (4%) in Shg levels in Strip-depleted cells. Genotypes: w,UAS-P35;ptcGAL4/UAS-cka.dsRNA at 18°C (A); w;ptcGAL4,UAS-GFP/+ at 25°C (B and E); w;ptcGAL4,UAS-GFP/UAS-strip.dsRNA at 25°C (C and F); w;ptcGAL4,UAS-GFP/UAS-cka.dsRNA at 25°C (D and G).