Validation of the AT1aR-Cre mouse line. a, Top, Wild-type AT1aR gene. Middle, To express both Cre-recombinase and zsGreen without interrupting AT1aR expression, 2A-Cre-2A zsgreen was introduced upstream of AT1aR 3′ UTR via homologous recombination. Bottom, F1 heterozygous mice were bred with a FLP-deleter strain, leading to the excision of the neomycin selection cassette. b–d, Projection images of the PVN collected from a mouse expressing both the AT1aR-Cre and the td-Tomato-stop-flox reporter gene depicting (b) AT1aR mRNA in cyan, (c) AT1aR-tdTomato-containing cells (AT1aR-Tom) in magenta, and (d) the merged image highlighting extensive overlap between the two markers. g–i, 10× images of coronal sections through the PVN of AT1aR-Tom reporter mice portraying (in magenta) the distribution of AT1aR-Tom throughout the PVN. White dashed lines outline the PVN unilaterally and the number on the bottom left of each image corresponds to the rostrocaudal distance from bregma. 3v, Third cerebral ventricle. Scale bars: b–d, 100 μm; e–i, 200 μm.