Fig. 1

Generation and validation of Nrg1-reporting mice. A Schematic diagram of the gene targeting strategy to insert the P2A-Cre cassette immediately before the stop codon of the Nrg1 locus, between exon 9 and 3’ untranslational region (3’UTR). The p2A peptide will be cleaved and two independent protein NRG1 and CRE will be expressed. B Southern blot screen for Nrg1^Cre/+ mice using Spel-digested genomic DNA and the Cre probe indicated in panel A. The targeted Nrg1 allele will yield a DNA fragment of 6.9 kb. C Southern blot screen for Nrg1^Cre/+ mice using Ndel-digested genomic DNA and the 5’ probe indicated in panel A. The wild-type and targeted Nrg1 allele will yield a DNA fragment of 5.5 kb and 6.6 kb, respectively. D The breeding strategy to get the Nrg1-reporting mice. The female Nrg1^Cre/+ mice were crossed with the male Rosa26^{LSL−tdTomato/+} (Ai14) mice to get the Nrg1^Cre/+; Rosa26^{LSL−tdTomato/+} mice, i.e., the Nrg1-reporting mice. E Double fluorescence in situ hybridization (dFISH) of tdTomato and Nrg1 mRNA in the striatum of Nrg1-reporting mice. The arrows indicate Nrg1-positive granule cells in the OB. Scale bar, 200 μm. F The enlarged image from the rectangle in panel E. Scale bar, 50 μm. G–H Similar NRG1 protein levels in the striatum between WT and Nrg1-reporting mice. G, representative western blots, the total lysates of striatum from WT and Nrg1-reporting mice were probed with anti-NRG1 and anti-GAPDH antibodies. H, quantification results. NS, not significant, n = 3, unpaired t-test