Extended Data Figure 6. TMT and bobcat urine activate CRH neurons and induce stress hormone increases.

a, Photographs of PVN sections from CRH-Cre mice crossed with Rosa-floxstop-GFP mice. Mice were exposed to water (H2O), rabbit urine, bobcat urine, or TMT and sections then costained with antibodies against GFP (green), to identify CRH neurons, and c-Fos (red), to detect activated neurons. Higher magnifications of boxed areas are shown at the right. Many CRH neurons (GFP+) expressed c-Fos after exposure to bobcat urine or TMT, but not rabbit urine. Scale bars = 100 μm (left) and 20 μm (right).

b, The percentage of PVN CRH (GFP+) neurons immunostained for c-Fos after odor exposure of CRH-Cre mice crossed with Rosa-floxstop-GFP mice. Exposure to bobcat urine or TMT, but not rabbit urine, increased the percentage of CRH neurons expressing c-Fos. (n=6 mice per condition. Error bars indicate SEM. ***p<0.001. One-way ANOVA with post-hoc Dunnett’s test.)

c, d, Plasma ACTH (c) or corticosterone (d) concentrations were measured in WT mice following odor exposure. Exposure of animals to TMT or bobcat urine increased blood levels of ACTH and corticosterone, whereas exposure to rabbit urine did not. (n=6 mice per condition. Error bars indicate SEM. *p<0.05, **p<0.01, ***p<0.001. One-way ANOVA with post-hoc Dunnett’s test (c) or Kruskal-Wallis test with post-hoc Dunn’s test (d).)