βarr2 is found in the axoneme and basal body of primary cilia.
(A) Confluent RPE1 cells were serum-starved for 24 hours, then fixed and stained for acetylated-tubulin (AT), which is highly enriched in primary cilia (axoneme) and for endogenous βarr2 using rARR antibody. (B) Z-stacks images of representative cells were deconvoluated as in Figure 2 and a 3D reconstruction of a representative cilium is shown. (C and D) RPE1 cells transfected with plasmids encoding for the βarr2-Cherry fusion (βarr2-Ch), were serum-starved for 24 hours after transfection, then fixed and stained for AT (C) or for the basal body marker pericentrin (D). In coloured images, βarr2 staining is in red, centrosome or cilia markers in green and nuclei stained with DAPI are in blue. (E) RPE1 cells transfected with plasmids encoding for Flag-tagged active form of smoothened (smo*) and the βarr2-Ch fusion were serum-starved for 24h after transfection, then fixed and stained for AT and smo*, using a rabbit polyclonal anti-Flag antibody. In coloured image, βarr2 staining is in red, AT in blue and smo* in green. Insets show higher magnifications of representative areas. Arrows stress basal bodies. Scale bars represent 5 µm. (F) Tissue sections from adult mouse kidney were stained for AT and for βarr2 using the rARR antibody. In coloured image, βarr2 staining is in red, AT in green and nuclei stained with DAPI in blue. The lumen of a representative tubule is indicated (Lu). Insets show higher magnifications of a representative ciliated tubular epithelial cell. Arrows stress AT positive structures.