Fig. 3. DdNek2 is a component of isolated centrosomes. (A) Western blot of cytosolic extract (CE; 2x10⁶ cells) and isolated centrosome (IC; 5x10⁷ cells) proteins separated on a 12.5% polyacrylamide gel (acrylamide:bisacrylamide, 200:1) and stained with anti-DdNek2 antibodies. (B,C), Immunofluorescence microscopy of isolated centrosomes spun down to coverslips, fixed with methanol and stained with rabbit anti-DdNek2 (B) and mouse anti-DdCP224 antibodies (C). Secondary antibodies were Alexa-568-labeled anti-rabbit IgG and Alexa-488-labeled anti-mouse IgG. Bar, 2 µm.