Characterization of Ebf2-EGFP+ cell-enriched genes in the developing mouse forebrain.

(A) Phase and fluorescent images of Ebf2-EGFP+ cells and Ebf2-EGFP- cells after FACS at Day 0 and Day 4 in vitro culture showing the purity and growth of sorted and unsorted cells (scale bar, 25 μm). (B) Sample to sample Pearson’s correlation (top-right quadrants) and pairwise comparison (bottom-left quadrants) of normalized gene expression between the two biological replicates and cell types. (C-D) qRT-PCR and endogenous expression patterns (In situ hybridization data from Allen Brain Atlas) to validate the RNA-seq results at different embryonic stages. CR neuron molecular markers as Calb2, Ebf2, Tbr1, and CR neuron related genes as Ebf1, Ebf3, Foxg1, and newly identified CR gene Nhlh2 were selected for validation. (E-F) qRT-PCR analysis of CR-specific genes in primary neural cell culture assay after lentiviral transduction of control (H1) or Ebf2-knockdown (E), or Foxg1-knockdown (F). Data represent mean ± SEM (n = 3 independent experiments, **P<0.01, *P<0.05, T test).

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