Fig. 4

Ythdf2^−/− NSPCs exhibit decreased proliferation and defects in natural differentiation in vitro. a Number of viable NSPCs at 0, 24, 72, and 120 h monitored by signal intensity of Presto Blue reagent. Proliferation rate was calculated by normalizing to wild type at 0 h. b mRNA expression levels of Ythdf2 during NSPC differentiation. Cells were collected at differentiation Day 0 (D0), 3, and 5. Isolated total RNAs were applied for RT-qPCR analysis. Actin was used as normalization control. c Immunostaining of Map2⁺ and Gfap⁺ cells differentiated from E14.5 neurospheres at D5 and D7. Nuclei were counterstained with DAPI. Scale bar indicates 20 μm. d Percentage of Map2 or Gfap positive cells. Error bars represent mean ± standard deviation, n = 3 biological repeats and 3 technical replicates. e Mean number of primary neurites per neuron (Map2⁺). n = 20 neurons for each biological repeat. f Mean length of the longest neurite of neurons (Map2⁺). n = 20 neurons for each biological repeat. *P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test