Fig. 2

Fig. 2. Knockdown of PCNT, KIF3B and TTC17 resulted in reduced number of cilia in cells. (a) Examination of the knockdown efficiency by siRNAs in human fibroblasts by real-time quantitative PCR assay using GAPDH as internal controls. Three independent experiments were performed and data represented as means ± standard deviation (SD). *** p < 0.0001, two-sided t-test. Cells transfected with non-specific scrambled siRNA were employed as negative controls and relative expression levels of PCNT, KIF3B and TTC17 to GAPDH in scrambled siRNA transfected cells were arbitrarily regarded as 1. (b) Effect of PCNT, KIF3B and TTC17 knockdown on cilia. Cells were transfected with siRNAs and serum-starved for 48 h, then immuno-stained for acetylated-α-tubulin (TUB; red) and pericentrin (PCNT, green). Nuclei were stained with DAPI (blue). Representative images of each of the siRNA transfection were shown. Ciliated cells (arrowheads) were abundant in scrambled siRNA transfected cells, whereas ciliated cells were scarce in siPCNT, siKIF3B and siTTC17 transfected cells. Ciliated cell and non-ciliated cells were magnified as shown as inset. (c) Percentage of ciliated cells in scrambled siRNA, siPCNT, siKIF3B and siTTC17 transfected cells was determined by counting ciliated cells and total number of cells. Two independent transfection experiments were performed for each siRNA, and over 100 cells were counted in each experiment. Data was shown as means±SD. *** p < 0.0001, one-way ANOVA, two-sided test. All scale bars = 50 µm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article).