Fig. 2: Altered Atoh1 levels in the hypoxic neonatal brain affected neuronal development.

a Heat map shows DEGs of the whole brain from P11.5 mice (control, n = 6; model, n = 6) (p < 0.05, Log₂FC > 1). b Bar graph shows TFs families in DEGs (control, n = 6; model, n = 6) (p < 0.05, Log₂FC > 1). c Volcano plot shows genes in the leading positions of DEGs (control, n = 6; model, n = 6) (p < 0.05, Log₂FC > 1.5). d Protein fractions in cytoplasm and cytoblast extracted from P11.5 mice brains were tested with Atoh1. Left panels show representative bands of Atoh1. β-actin and H3 were used as internal controls of cytoplasm and cytoblast, respectively. Bar graphs on the left show percentage changes of proteins in model mice relative to control. Bar graph on the right shows ratio of Atoh1 level in cytoblast relative to cytoplasm. (control, n = 3; model, n = 3) (For statistics, see Table S9). e Staining of cerebellar lobule III by antibodies against Atoh1 and NeuN in P11.5 mice. Cb, cerebellum. Scale bars: 20 μm. Objective 40x. Bar graph shows MFI of Atoh1 staining (control, n = 6; model, n = 6) (t = −3.89, **p = 0.0030, df = 10) (1.00 ± 0.033 vs 1.20 ± 0.040). f Protein fractions from P11.5 mice brains of control and model mice were tested with TrkB, BDNF, SYP, Sema4F, Atoh1, pErk1/2, and Erk1/2. Left panels show representative bands of proteins mentioned above. α-tubulin was used as an internal control. (NC, negative control; OE, over expression). Bar graphs show percentage changes of proteins (control + NC, n = 3; model + NC, n = 3; model + OE-Atoh1, n = 3) (For statistics, see Table S10). g Sholl analysis of cultured primary neurons stained of SYP. Left panels show representative staining of SYP and intersection of dendrites with concentric circles. Line chart shows the intersection numbers of dendrites and concentric circles. Scale bars: 20 μm. Objective 63x. (All groups, n = 6 batches for each group, n = 2 cells for each batch) (For statistics, see Table S11). All data were shown as mean ± SEM. *p < 0.05, **p < 0.01.