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Fig. 2 | Molecular Autism

Fig. 2

From: Impaired neurogenesis and neural progenitor fate choice in a human stem cell model of SETBP1 disorder

Fig. 2

Neuronal differentiation is affected by loss of SETBP1 A Schematic representation of hESC cortical differentiation protocol. B Expression of NPC markers at day 18. Cultures were immunostained for N-cadherin (red) and DAPI (blue) showing the organization and size of the neural rosettes, PAX6 (green), OTX2 (red), NESTIN (NES, green), SOX2 (red) and FOXG1 (red). Dapi was used to label all nuclei. Scale bar: 100uM. C Quantitative data of marker expression presented as mean ± s.e.m for each genotype with a minimum of two independent experiments carried out per line and minimum of four per genotype (WT = 7, HET1 = 2, HET2 = 2, Homo1 = 6 and Homo 2 = 2). One-way ANOVA test with Bonferroni Post Hoc or Kruskal–Wallis non-parametric test, SOX2 P = 0.890, NES P = 0.499, OTX2 P = 0.606, FOXG1 P = 2.159E−06 (Post Hoc WT vs. SETBP1-/-, p = 0.000032; WT vs. SETBP1+/- , p = 0.331; SETBP1+/-  vs. SETBP1-/-, p = 3.77E−06), PAX6 P = 0.018 (Post Hoc WT vs. SETBP1-/-, p = 0.035; WT vs. SETBP1+/- , p = 1; SETBP1+/- vs. -/-, p = 0.052). (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). D Immunostaining of cortical layer markers TBR1 (layer VI) and CTIP2 (layers V–VI) at days 30, 40 and 50, respectively, and SATB2 (layers II–III) at day 50. Images representative of several independent experiments for each genotype E Quantitative analysis of the above. Data presented as mean ± s.e.m for each genotype with a minimum of two independent experiments carried out per line (WT = 5, Het1 = 2, 2 = 2, Homo1 = 3, and Homo2 = 2). One-way ANOVA test, Bonferroni Post Hoc; TBR1 p = 0.615 for day 30, 0.863 for day 40, and 0.585 for day 50; CTIP2 P = 0.004 for day 30 (Post Hoc WT vs. SETBP1-/-, p = 0.006; WT vs. SETBP1+/-, p = 1; SETBP1+/- vs. -/-, p = 0.007), 0.022 for day 40 (Post Hoc WT vs. SETBP1-/-, p = 0.05; WT vs. SETBP1+/-, p = 1; SETBP1+/- vs. -/-, p = 0.05), and 0.042 for day 50 (Post Hoc WT vs. SETBP1-/-, p = 0.0071; WT vs. SETBP1+/-, p = 1; SETBP1+/- vs. -/-, p = 0.182); SATB2 P = 0.491 for day 40, and 0.348 for day 50 (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). Scale bar: 100uM

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