Fig. 2: Non-reduced Igfbp2 is necessary for NSC lineage commitment.

a, b Second-passage WT and Ncf1^–/– NSCs were treated with mouse recombinant insulin-like growth factor-binding protein 2 (Igfbp2) or H₂O₂ 24–28 h after plating. a Photomicrographs of NSCs 5 days after treatment with Igfbp2 (60 ng ml^–1) or H₂O₂ (10 µM). b Quantification of neurospheres ≥60 µm in diameter (n = 4 donors). Western blot below the bar graph shows Igfbp2 levels in the medium 5 days post-treatment. c Photomicrographs of NSCs 8 days after treatment with 100 ng ml^–1 native Igfbp2, H₂O₂-treated (Oxi), DTT-treated (Red), or DTT then H₂O₂ sequentially treated (Red → Oxi) Igfbp2. d Quantification of neurospheres ≥60 µm in diameter (n = 6 donors). e, f Igfbp2^–/– NSCs were transfected with empty or Igfbp2 expressing plasmid 24 h after plating. e Photomicrographs of NSCs 11 days after transfection. f Quantification of neurospheres ≥60 µm in diameter (n = 6 donors). g, h Second passage WT and Igfbp2^–/– NSCs were treated with H₂O₂ 24 h after plating. g Photomicrographs of NSCs 10 days after H₂O₂ (10 µM) treatment. h Quantification of neurospheres ≥60 µm in diameter (n = 6 donors). Two-tailed students t-test (f), Two-way ANOVA (b), and One-way ANOVA (d, h), followed by Bonferroni multiple comparison posttest. Error bars indicate s.e.m. Scale: 200 µm (a, c, e) and 300 µm (g). Source data are provided as a Source data file.