Mice were anesthetized (350 mg/kg chloral hydrate, i.p.) and transcardially perfused with 4% paraformaldehyde in 0.1M sodium borate buffer at 4°C, as described previously46. Brains were post-fixed for 2h at 4°C and then incubated in 0.05 M potassium-PBS (K-PBS) containing 15% (w/v) sucrose 4°C for 12–16 hr. Brains were sectioned on a sliding microtome (25 μM to 30 μM), collected in equally-spaced series and stored in cryoprotectant (20% glycerol and 30% ethylene glycol in 0.1 M phosphate buffer) at −20°C. Immunohistochemistry was conducted on free-floating sections by the avidin-biotin-complex method using the chromogen, diamino-benzidine (Vector Labs). For P-STAT3 (Y705) immunohistochemistry (IHC), sections were pre-treated with 1% H2O2 in 1% NaOH for 10 min, 0.3% glycine in K-PBS for 10 min, and 0.03% SDS in K-PBS for 10 min 47. For TORC1, CART, and KISS1 IHC, sections were pre-treated with 0.3% H2O2 for 10 min and 0.3% glycine, 0.3% Triton X-100 in K-PBS for 10 min. Brain sections were blocked in a K-PBS solution containing 1% Probumin (Millipore), 1% normal donkey serum (Jackson ImmunoResearch) and 0.03% Triton-X-100. Sections were incubated overnight
