Fig. 4: Ncf1/Igfbp2 axis promotes NSC lineage-commitment through repression of NSC DNA repair pathways.

a, b Photomicrographs (a) and quantification (b) of neurospheres ≥60 µm in diameter 7 days after transfection of Ncf1^–/– NSCs with siRNA targeting Fanca, Fancd2 or Rad51 or negative control (n = 6 donors). c, d Western blot analysis (c) and quantification (d) of γ-H2ax levels (normalized to Gapdh) in NSC lysates 3 days after Igfbp2 treatment (n = 7 donors from two different experiments). e, f Western blot (e) and quantification (f) of γ-H2ax levels (relative to Gapdh) in cell lysates 3 days after transfection (n = 3 donors). The samples were derived from the same experiment and blots were processed in parallel. kD = kilodalton. g–i WT NSCs were transduced with Peggy Back transposase and transposon to overexpress GFP or Fanca. g Photomicrographs and h quantification of neurospheres ≥60 µm in diameter ten days after plating P4 NSCs overexpressing GFP or Fanca (n = 6 donors). i qPCR of Fanca mRNA 3 days after plating P5 WT NSCs overexpressing GFP or Fanca (n = 6 and 5 donors in GFP and Fanca OE NSCs respectively). ΔΔCT: the difference in threshold cycles normalized to β-Actin. Kruskal–Wallis test and FDR method of Benjamini and Hochberg multiple comparison posttest (d), One-way ANOVA and FDR method of Benjamini and Hochberg (b) or Bonferroni multiple comparison posttest for marked comparisons in (f) and two-tailed Mann–Whitney U test (h, i). Error bars indicate s.e.m. Scale: 120 and 250 µm in (a) and (g) respectively. Source data are provided as a Source data file.