Catalogue sc 52 g

For all immunohistochemical experiments, mice were anesthetized (Beuthanasia, 320 mg/kg delivered ip) and intracardially perfused with 0.1 M PBS followed by 4% paraformaldehyde. Brains were then extracted, post-fixed in 4% paraformaldehyde overnight, and cryoprotected in 0.1 M PBS containing 20% sucrose until the brains sunk in the sucrose solution. Coronal cryostat sections 30-µm thick were collected and every third section of the PBN and BNST, or every fourth section of the CeA, were processed for immunolabeling and quantification. For co-labeling of GFP and Fos, sections were incubated for 16 h at room temperature in chicken anti-GFP (1:10000, catalogue # ab13970, lot #GR236651-4, Abcam) and goat anti-Fos (1:700, catalogue # sc-52-G, lot #F1615, Santa Cruz Biotechnology). Because we ran out of our original Fos antibody, tissue from Figure 4 was stained with goat anti-Fos from a different lot (1:700, catalogue # sc-52-G, lot #F1616 Santa Cruz Biotechnology). The sections were then washed and incubated for 2 h at room temperature in Alexa488-conjugated donkey anti-chicken (1:400, Jackson ImmunoResearch) and CY5-conjugated donkey anti-goat (1:400, Jackson ImmunoResearch).