Fig. 2
From: Knocking out ARL13B completely abolishes primary ciliogenesis in cell lines
ARL13B-KO HEK293T cells have no cilia. (a) The genotypes of ARL13B-KO HEK293T cell lines, sg1#5 and sg2#5. See the legend of Fig. 1a. (b–e) Endogenous ARL13B is undetectable by Western blot and immunofluorescence imaging, and no cilia could be found in sg1#5 and sg2#5 cells. In (b), anti-GAPDH was used for loading control. Non-specific bands are indicated by asterisks (*). Molecular weight markers are labeled to the right of the immunoblot. See the legend of Fig. 1b. (c) Fluorescence imaging of endogenous ARL13B and acetylated tubulin in parental and KO HEK293T cells. Cells were fixed and co-immunostained by antibodies against indicated cilium markers before being subjected to wide-field microscopy. (d) Quantification of the ciliogenesis percentage in parental and KO HEK293T cells. Images acquired as described in (c) were analyzed. n = 3 independent experiments with ≥ 100 cells analyzed in each case. The error bar represents the mean ± standard deviation. (e) Fluorescence imaging of endogenous glutamylated tubulin and pericentrin in parental and KO HEK293T cells, similar to (c). In (c and e), the boxed region is enlarged at the lower left corner, and the scale bar represents 10 μm.
