Fig. 4.

GFP-Ipk1 is enriched in centrosomes and at basal bodies of cilia. Localization of GFP-Ipk1 in HeLa (A–D) and LLC-PK1 Cl4 (E–L) cells is shown. Cells transfected with pEGFPc1-Ipk1 were fixed in 3% paraformaldehyde and imaged for GFP fluorescence. Coimmunostaining with anti-γ-tubulin (B) and anti-pericentrin (C and K) antibodies detected centrosomes. Anti-acetylated-tubulin antibodies detected cilia (F and J), and TO-PRO3 staining was for nuclear DNA (G). Overlays show colocalization of GFP Ipk1 with γ-tubulin/pericentrin in HeLa cells (D), and with pericentrin and/or at the base of the cilium in LLC-PK1-Cl4 cells (H and L). White arrowheads in A, E, and I indicate GFP-Ipk1 foci. (Scale bars, 5 μm.)