Extended Data Fig. 3: AKNA dissociates from the centrosome during mitosis and upon increased phosphorylation.

a, Immunofluorescence of AKNA in primary E14 cortical cells at different phases of the cell cycle, showing the lack of AKNA immunofluorescence at the centrosome during mitosis (n = 3 independent experiments). b, Western blot of AKNA in synchronized A20 cells, showing that AKNA protein is not degraded during mitosis (as indicated by the presence of phospho-histone H3) (n = 2 independent experiments). c, Representative micrographs of AKNA and PCNT immunofluorescence in E14 primary cortical cells treated with 500 nM okadaic acid (OA). Note that AKNA immunofluorescence is observed at centrosomes at 0 h but is undetectable in most cells 3 h after treatment, which shows that the centrosomal localization of AKNA is phosphorylation-dependent (n = 3 independent experiments). d, Western blot of lysates of OA-treated cells shows that phosphorylation—here caused by protein phosphatase inhibition—shifts that AKNA band on SDS–PAGE, and subsequently leads to protein degradation as observed in lysates of cells 5 h after OA washout (n = 3 independent experiments). Scale bars, 5 μm (a), 10 μm (c).