Fig. 6.

Ars2 targets essential neural regulators. (A) Functional annotation of Ars2-bound regions in proliferating NSCs. (B) Ars2 and SOX2 enrichment profiles at representative NSC regulators (e.g. Hes5 and Notch1) and glial regulator Olig2. (C) Selected Ars2 peaks were validated by ChIP-qPCR (red bars in panel B indicate amplicon sites) with our independent Ars2 antibody; i.e. different from the one used for Ars2 ChIP-seq. SOX2 ChIP-qPCR indicates genomic colocalization. Experimental material was treated with RNase before PFA fixation and ChIP. Data are mean±s.e.m. (D) Schematic of Cre-virus infection strategy and qPCR measurements of NSC genes (e.g. Hes1, Hes5 and Sox2) from primary Ars2^flox/flox NSCs 48 h after Cre-virus infection. Data are mean±s.e.m. Hes1 P=0.0053; Hes5 P=0.0140; Sox2 P=0.0001; two-tailed t-test applied; data are mean±s.e.m. (E) Strategy to knockout Ars2 in embryonic ventral NSCs using Olig2-Cre. Red region in the diagram indicates Olig2-Cre-expressing area in embryonic ventral forebrain. (F) E15.5 Olig2-Cre; Ars2^fl/+ (ctrl) and Olig2-Cre; Ars2^fl/fl medial ganglionic eminence (MGE) region stained for Ki67 (red), BrdU (green) and DAPI (blue). Arrowhead indicates that the thickness of MGE, the ventral progenitor region, is reduced in mutants. (G) E15.5 Olig2-Cre; Ars2^fl/fl ventral VZ stained for SOX2 (red), Ars2 (green) and DAPI (blue). Arrowheads indicate that cells without Ars2 are specifically negative for SOX2. Panels on the right show magnification of the boxed areas on the left. Scale bar: 100 µm in F; 50 μm in G.