Fig 1: Subcellular localization of Cep215 in P19 cells under differentiation. (a) Experimental scheme of glial differentiation of P19 cells. The cells were treated with retinoic acid (RA) for 4 days to induce embryoid body (EB) and cultured for up to 17 days. Neurogenesis occurs at the early stage of differentiation (D4-8) whereas gliogenesis occurs at the late stage of differentiation (D9-13). (b) P19 cells were treated with RA for differentiation and subjected to immunoblot analysis with antibodies specific to Nf68, Gfap and Gapdh. (c) The P19 cells at D17 were subjected to coimmunostaining analysis with antibodies specific to Cep215 (red) along with Tuj1 or Gfap (green). (d–f) Undifferentiated (UD) and differentiated (D12 and D15) P19 cells were subjected to immunoblot (d) and coimmunostaining (e) analyses with antibodies specific to Cep215, Gapdh and Gfap. (f) Intensities of the centrosomal Cep215 signals were measured. In case of D12 and D15, only Gfap-expressing cells were subjected to analysis. Greater than 90 cells per experimental group were estimated in three independent experiments. The statistical significance was analyzed using one-way ANOVA. *P < 0.05. (c, e) Nuclei were stained with DAPI (blue). Scale bars, 10 µm.