Divergent roles of the Wnt/PCP Formin Daam1 in renal ciliogenesis
Fig 3
The sh-daam1 cilia phenotype can be rescued with wild-type Daam1 but not Daam1(I698A).
Stable MDCKII cells infected with either sh-daam1 or a control sh-scrambled construct were transiently transfected with constructs that express either GFP, GFP-Daam1, or GFP-Daam1(I698A) constructs. Cells were grown on transwell filters and stained with acetylated α-Tubulin antibody (acTubulin) to label primary cilia (green), DAPI to label nuclei (blue), and phalloidin to label F-actin (Magemta). Confocal imaging was used to analyze the effects upon primary ciliogenesis. A) Representitive images of ciliated cells stained with acetylated tubulin (green), dapi (blue) and phalloidin (magenta). Scale bars equal to 50 μm. B) Quantification of the amount of ciliation. Error bars are shown as ± SD. Black dots indicate individual values for each image quantified. * indicates p < 0.05 as compared to sh-scrambled using two tailed t test.