Fig. 6.

Acetylated tubulin forms a complex with S-palmitoylated Arl13b in cilia. A–C: Metabolic labeling of HEK293T cells with PAz, followed by click chemistry–mediated addition of fluorophore (Cy5) and immunoprecipitation using anti-Arl13b rabbit IgG. Immunoblot shows coimmunoprecipitated acetylated tubulin, the signal of which is reduced by 2BPA (arrow in B). Quantitation is shown in C. N = 3. P < 0.01. D: PLA for complex of Arl13b with acetylated tubulin (signal in red, arrow) in HEK293T cells. Acetylated tubulin labeling is shown in green and Arl13b in blue. E, F: PLA for S-palmitoylation of Arl13b in HEK cells (E) and ependymal cells (F). Arrows point at S-palmitoylated Arl13b (red) in cilia. Acetylated tubulin labeling is shown in green. αArl, immunoprecipitation using anti-Arl13b rabbit IgG (immunolabeled Arl13b is pseudocolored in green, and palmitoylation is pseudocolored in red); 2BPA, 2-bromo palmitic acid; IgG, control immunoprecipitation using nonspecific rabbit IgG; In, input; PAz, palmitic azide; PLA, proximity ligation assay.