Chd5 deficiency in NSCs leads to premature activation.

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Chd5 deficiency in NSCs leads to premature activation. Chd5 deficiency in NSCs leads to premature activation. (A) Representative phase contrast images of wild-type (+/+) and Chd5-deficient (Chd5−/−) postnatal day 1 (P1) NSCs grown as adherent cultures (upper images) and neurosphere cultures (lower images). Scale bars = 100 μm (upper images) and 200 μm (lower images). (B) Quantification of the size of wild-type and Chd5-deficient NSC neurospheres expressing empty vector (EV) or Chd5 cDNA (Chd5). Data are represented as mean ± SD (n = 3). ns, no significance; **<0.01; ***<0.001; Tukey's multiple comparison test. (C) Flow cytometry of EdU-incorporated populations of wild-type and Chd5-deficient NSCs. Data are represented as mean ± SD (n = 4). ***<0.001; unpaired t test. (D, E) Flow cytometry of Egfr-positive and Cd133-positive populations in wild-type and Chd5-deficient NSCs. Data are represented as mean ± SD (n = 3). ns, no significance; *<0.05; unpaired t test. (F, G) Immunofluorescent images and Western blots of wild-type and Chd5-deficient NSCs, assessed for Chd5 (red), nestin (green), and DAPI nuclear signal (blue) (left panels), or assessed for Pax6, nestin, and β-actin (actin) expression (right panels). Scale bar = 50 μm. (H, I) Representative immunofluorescent images of wild-type and Chd5-deficient NSCs, assessed for vimentin (red), nestin (green), and DAPI nuclear signal (blue), and corresponding distribution of mean signal intensities over pixels of nestin and vimentin expression in randomly selected fields (4–5 fields per sample, n = 2–3). Scale bar = 50 μm. Dong-Woo Hwang et al. LSA 2018;1:e201700016 © 2018 Hwang et al.