Fig. 1
MPP9 represents the ring-like structures at the distal ends of the centrioles. a Immunofluorescence of MPP9 (green) and Centrin-3 (red, upper) or C-Nap1 (red, lower) in U2OS cells. b Immunofluorescence of MPP9 (green) and acetylated-tubulin (Acet-Tub, red) in mitotic HeLa cells. DNA was stained with DAPI (white). c Immunoblots of MPP9 during the cell cycle in hTERT RPE-1 cells. Tubulin was used as a loading control. Relative amounts of MPP9 were quantified and normalized to tubulin. d 3D-SIM images of U2OS cells triple-immunostained with antibodies against MPP9 (green), CP110 (purple), and Centrin-3 (red). Arrows show distal ends of the centrioles. e 3D-SIM images of MPP9-GFP (green) overexpressing U2OS cells co-immunostained with antibodies against CP110 (red) and C-Nap1 (purple). Arrows show proximal ends of the centrioles. f–i 3D-SIM images of U2OS cells co-immunostained with antibodies against MPP9 (f, green) and Acet-Tub (f, red) or MPP9 (h, red) and CEP164 (h, green). g, i are the intensity plots of the rings from f, h, respectively. j Immuno-electron microscopy images. U2OS cells were labeled with anti-MPP9 antibodies followed by anti-rabbit IgG-gold (10 nm) secondary antibodies. Green, gold particles. Schematics of immuno-electron microscopy images are shown. k Schematic of MPP9 localization at centrosomes. Scale bars: 5 μm (b); 1 μm (a); 500 nm (d, e)
