Extended Data Fig. 4: Temporal and sub-type-specific regulation of AKNA in the developing telencephalon.

a–c, Micrographs showing that PCNT⁺ centrosomes lack AKNA in the cerebral cortex at E9 (a, n = 2 experiments) and E18 (b, n = 2 experiments), whereas AKNA is enriched in the ventricular zone and—specifically—the SVZ at E14 in the ganglionic eminence (c, n > 10 experiments) and the cerebral cortex (Fig. 1). d, Distribution of AKNA⁺ centrosomes in the E14 cortex (n = 3 independent experiments). e–g, Plots showing the percentage of AKNA⁺ centrosomes in E14 and E13 cerebral cortex regions as indicated (e, f, n = 3 independent experiments), and in dissociated primary E14 cortical cells (g), which reveals that cells with AKNA⁺ centrosomes are mostly differentiating NSCs (PAX6⁺TBR2⁺) and PAX6⁻TBR2⁺ basal progenitors (n = 3 independent experiments). EM, electron microscopy; IF, immunofluorescence. h, Micrographs of cells isolated from E14 cerebral cortex by FACS using prominin 1, and stained for PAX6 (red arrows) and TBR2 (blue arrows). This shows that double-positive (differentiating) NSCs have AKNA⁺ centrosomes, whereas PAX6⁺TBR2⁻ (proliferating) NSCs do not (n = 2 experiments). Data in d–g are presented as mean ± s.e.m. Scale bars, 10 μm (a, c, h), 20 μm (b).

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