HSV-1 Infection Model in CD-1 Mice

Six-week male CD-1 (ICR) mice were purchased from Shanghai Laboratory Animals Center. For HSV-1 infection, mice were anesthetized by intraperitoneal injection of 0.4 ml of a mixture consist of 500 ug/ml xylazine hydrochloride (Sigma, X1251) plus 4 mg/ml pentobarbital sodium (Solarbio) in sterile saline. Then 2 x 10⁵ pfu of virus was added onto each scarified cornea in 3 μl of PBS. For TG acquisition, mice were euthanized by cervical dislocation while under anesthesia with isoflurane and TG were collected into the lysis buffer and placed on dry ice before RNA extraction and placed into -80°C as described previously [59 (link)]. For western blot analysis, we homogenized TG in lysis buffer (Solarbio, R0020) and extracted protein according to the manufacturer’s protocol. For determination of viral titers in TG, TG were homogenized in cell culture media for titer determination. For eye swab collection, mice were anesthetized in an induction chamber with isoflurane (3% in oxygen 0.5 ml/min) using a V1 Table Top anesthesia machine (Colonial Medical Supply). Both eyes of each mouse were swabbed with cotton-tipped applicators, suspended in 1 ml of cell culture medium.