FIG. 1.

Isolation of Dgcr8^−/− NSCs from conditionally Dgcr8-disrupted mouse embryonic brain. (A, B) Bright field images of NSCs isolated from mouse E13.5 embryonic brain. (A) Dgcr8^+/− and (B) Dgcr8^−/− NSCs. Scale bars, 50 μm. (C) PCR genotyping of NSCs. Shown are genotyping results of a wild-type control (mouse tail tip fibroblasts), two independent clones of Dgcr8^+/− NSCs, and two independent clones of Dgcr8^−/− NSCs. Arrow, wild-type Dgcr8 allele (Dgcr8⁺); arrowhead, Dgcr8 mutant allele (Dgcr8⁻). (D) qRT-PCR analyses of Dgcr8 (left) and Neurog2 (right) in Dgcr8^+/− and Dgcr8^−/− NSCs. Data were normalized to the mRNA levels of β-actin gene Actb. n = 3 independent biological repeats. Error bar, SD. (E, F) Immunostaining of NSC markers SOX2 (green) and NESTIN (red) in (E) Dgcr8^+/− and (F) Dgcr8^−/− NSCs. Cell nuclei were counterstained with DAPI (blue). Scale bars, 50 μm. NSC, neural stem cell. Color images available online at www.liebertpub.com/scd