Fig. 4

Akt is required for the maintenance of cilia in the ventral epithelia. a Gene expression levels of cilia specific markers at days 10, 20 and 30 after Smed-AKT(RNAi) relative to the control. Analysis of markers corresponding to intraflagellar transport machinery (IFT88) and flagellar beating (IC2, LC1, DNAHβ-1 and LRRC50) reveal a marked reduction of gene expression as early as day 10 and it is further reduced by day 30 after first injection in Smed-AKT(RNAi) animals. Gene expressions are all relative to the internal control, the ubiquitously expressed clone H.55.12e. Graphs represent mean ± s.e.m. of triplicated samples of two or more biological replicates with at least ten animals per experiment. Significance **** <0.0001 is determined through two way-ANOVA. b Images show control (top) and Smed-Akt (RNAi) (bottom) at day 30 after first dsRNA injection. Images represent depth-coded maximum projections of confocal z-sections in the posterior specific ventral cilia and proximal structures stained with Ac-Tubulin antibody. Superficial structures were labeled in magenta and structures near the dorsal epithelial layer appear in orange (color code scale on the left). Magnified images (on the right) display the effects of Smed-AKT(RNAi) on ciliated structures (cilia in ventral epithelia is represented in magenta and sections of the protonephridia in green and blue). Box with dotted lines outline regions of cilia in ventral epithelia, notice the scattered magenta signal in the experimental group. Confocal z-sections were taken on three or more animals of two independent experiments. Depth-coded projections were obtained using ImageJ software. Scale bar 100μm