Disrupted dynamics of early stages of differentiation perturbs neural cell fate specification. Disrupted dynamics of early stages of differentiation perturbs.

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Disrupted dynamics of early stages of differentiation perturbs neural cell fate specification. Disrupted dynamics of early stages of differentiation perturbs neural cell fate specification. (A) Representative immunofluorescent images of wild-type (+/+) and Chd5-deficient (Chd5−/−) NSCs that were undifferentiated (0 h) or differentiated (for 3, 6, 12, and 24 h) and assessed for vimentin (red), nestin (green), and DAPI nuclear signal (blue). Scale bar = 50 μm. (B, C) Distribution and comparison of mean signal intensities over pixels of nestin expression in randomly selected fields (4–5 fields per sample) of wild-type (upper panels) and Chd5-deficient (lower panels) NSCs at the time points, indicated in representative images in Fig S1A. Data are represented as mean ± SD (n = 2). ns, no significance; **<0.01; ****<0.0001; Tukey’s multiple comparisons test. (D) Temporal representation of mean signal intensities over pixels of nestin expression of wild-type (blue) and Chd5-deficient (gray) NSCs, as plotted in Fig S1B and C. Data are represented as mean ± SD (n = 2). (E) Immunofluorescent images of wild-type and Chd5-deficient NSCs that were differentiated for 48 h and assessed for Gfap (red), Map2 (green), and DAPI nuclear signal (blue). Scale bar = 50 μm. (F) Western blots of P1 neonatal wild-type and Chd5-deficient cortices and assessed for Chd5, Map2, and β-actin (actin) expression. Arrow indicates the band with a predicted molecular weight. (G) Western blots of undifferentiated wild-type and Chd5-deficient NSCs (0 h post-differentiation) and differentiated cells (3-h post-differentiation), assessed for Tbr2, Tbr1, and β-actin (actin) expression. Arrow indicates the band with a predicted molecular weight. Dong-Woo Hwang et al. LSA 2018;1:e201700016 © 2018 Hwang et al.