2.8. Immunohistochemistry

Tissues were collected as previously described [26]. Tissue was stained overnight with primary antisera (Table 1) before incubation with appropriate secondary antisera (Table 1). Slides were imaged using an Axioscan Z1 slide scanner (Zeiss) and confocal microscope (Leica TCS SP8 X). For c-Fos studies, 2–6 sections per mouse at the level of the hypothalamus (bregma −1.58 to −2.3 mm) and hindbrain (bregma −7.2 to −7.76 mm) were counted bilaterally and averaged across sections for each mouse. Images were analysed in ImageJ.

Antibodies.