Vero, BHK, and Raw264.7 Cell Culture Protocols

The Vero cells (African green monkey kidney epithelial cells) and the BHK 21 cells (Baby hamster Kidney epithelial cells) were maintained in the Dulbecco’s modified Eagle’s medium (DMEM; PAN Biotech, Aidenbach, Germany) supplemented with 5% Fetal Bovine Serum (FBS, PAN Biotech, Aidenbach, Germany), 0.1% Gentamycin and 1% Penicillin–Streptomycin (PAN Biotech, Aidenbach, Germany). The Raw264.7 cells (mouse monocyte/macrophage cell line), was maintained in RPMI-1640 (Himedia Laboratories Pvt. Ltd, Mumbai, India) supplemented with 2.0 mM L-glutamine, Penicillin 100 U/mL, Streptomycin 0.1 mg/mL and 10% Fetal bovine serum (FBS; PAN Biotech, Germany) at 37 °C under a humidified incubator with 5% CO₂. The HSV-1 virus strain KOS with GenBank accession Number JQ673480.1 [15 (link)] was kindly gifted by Dr. Roger Everett, Glasgow University, Scotland. MBZM-N-IBT was synthesized by our group [13 (link)] and acyclovir was procured from Sigma (Sigma, USA). The anti-ICP8 (ab20194), anti-gC (ab6509) monoclonal antibodies and GAPDH antibody were procured from Abcam (Cambridge, UK) and Abgenex Pvt. Ltd. (Bhubaneswar, India) respectively.