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jws-dvdy.21072.img1.tif1563K Supplementary Figure 1 : Sox2 and MAP2 immunohistochemistry demarcate neural progenitors and their neuronal progeny respectively.A)A section of E10 developing cerebral cortex shows the presence of Sox2 positive neural progenitors in the ventricular zone.B)A section of cerebral cortex at birth shows the MAP2 positive neurons. Cortex (Cx), Lateral ventricle (LV), Ventricular zone (VZ).C)The undifferentiated Sox2 positive P19 cells.D)The MAP2 positive P19 neurons after 8 days in vitro. Scale bar:C(15 ?m),D(20 ?m),EandF(10 ?m).
jws-dvdy.21072.img2.tif2116K Supplementary Figure 2 : numb4-EGFP is stably expressed in P19 cells.Using lipofectamine, P19 cells maintain numb4-EGFP expression in undifferentiated adherent cultures(A)and aggregates treated with RA for 4 days(B) .C)The corresponding phase contrast image of B.D-E)P19 neurons are present at days 8(D)and 12(E)in vitro, based on neurite growth. Scale bar: 20 ?m.
jws-dvdy.21072.img3.tif1111K Supplementary Figure 3 : MASH1 immunoreactivity is detected in P19 EGFP clone upon RA induction. A)Undifferentiated P19 cells do not express MASH1 prior to RA treatment.C)MASH1 is expressed in cell aggregates upon RA induction.BandD)The corresponding phase contrast images of A and C, respectively. Scale bar: 30 ?m.

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