Immunohistochemical Analyses of FoxP2 and Xpress

For histological analyses, animals were sacrificed 3–5 days following HSV injection then perfused with warm saline followed by ice cold 4% paraformaldehyde in 0.1 M phosphate buffer. Tissue was cryosectioned at 20 µM, thaw-mounted onto glass microscope slides, and stored at −80°C until use. Thawed sections were incubated overnight with goat-anti-FoxP2 (1:500; Abcam, Cambridge, UK; [Thompson et al., 2013 (link)]) and mouse-anti-Xpress (1:500; ThermoFisher Scientific, Waltham, MA). AlexaFluor 546 donkey-anti-goat (1:500) and AlexaFluor 405 donkey-anti-mouse (1:250) secondary antibodies were used to generate anti-FoxP2 and anti-Xpress signals, respectively. Sections were visualized using a Zeiss (Oberkochen, Germany) LSM 800 confocal microscope and processed using NIH ImageJ (Schneider et al., 2012 (link)).