Donkey anti rabbit antibody conjugated to cy3

vmPFC–amygdala: ChR2 mice received blue light in the amygdala (10 Hz, 5-ms pulses, 10 mW power) for 10 min. Animals were perfused 90 min after receiving optical stimulation. Following perfusion, brain slices were obtained as described above in ‘Histology’. c-Fos staining was performed as described previously^{46 (link)}. In brief, coronal sections containing the amygdala were washed in PBS (three 10-min washes), and were then incubated for 1 h in blocking solution with 0.2% Triton-X-100 and 2% normal donkey serum. Sections were then incubated overnight with anti c-Fos primary antibody (rabbit anti c-Fos 1:500, Cell Signaling Technology, cat. no. 2250S). The next day the sections were washed in PBS (three 10-min washes) and incubated at 1 h at room temperature with the secondary antibody at 1:500 dilution (donkey anti-rabbit antibody conjugated to Cy3, Jackson Laboratories). Slices were washed three times in PBS and mounted on slides with PVA-Dabco (Sigma). c-Fos counting was performed blinded to treatment group on a Leica TCS SP5 scanning laser confocal microscope. To stain cells against GABA the same procedure was employed, but this time using anti-GABA antibody (Sigma, cat. no. A2052).