Figure 2.

Mutation of the EXOC5 ciliary targeting sequence inhibits ciliogenesis. A, control, EXOC5-OE, Exoc5 KD, and EXOC5CTS-m (mut) stable cell lines were grown on Transwell filters to confluency. All cell lines reached confluency at the same time, as determined by a fluid maintenance test (36). The Transwell filters were fixed, and the cells were stained with nuclear DAPI (blue) and acetylated α-tubulin antibody against ciliary axonemes (red). B, higher magnification image of control, EXOC5 OE, Exoc5 KD, and EXOC5CTS-m MDCK cells stained with DAPI (blue) and acetylated α-tubulin antibody (red). C, quantitation of the percent of cells that have primary cilia shows that EXOC5 OE cells are more ciliated, whereas Exoc5 KD and EXOC5CTS-m are less ciliated, than control MDCK cells. D, quantitation of ciliary length shows longer cilia in EXOC5 OE, compared with control, MDCK cells. n = 700 cells counted for each cell line, with the experiment repeated three times.