Dulbecco s modified eagle medium dmem 1

All experiments were performed using human immortalized foreskin fibroblasts (HFFs) that were immortalized by hTERT transfection (kindly provided from the Sara Selig lab). All viruses were grown and tittered on African green monkey kidney epithelial cells (Vero cells; ATCC CCL-81) or, in the case of ICP0 mutant viruses, human female osteosarcoma cells (U2OS cells; ATCC HTB-96). 293FT cells, a variant of the human kidney cell line 293, expressing simian virus 40 (SV40) large T antigen (provided by the Chen Luxenburg lab), were used for obtaining the VP16-expressing lentivirus. All cells were grown with Dulbecco’s modified Eagle medium (DMEM ×1; Gibco), supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin and streptomycin (10,000 units/mL and 10 mg/mL, respectively; Biological Industries, Israel).

All experiments were performed using human immortalized foreskin fibroblasts (HFFs) that were immortalized by hTERT transfection (kindly provided from the Sara Selig lab). All viruses were grown and tittered on African green monkey kidney epithelial cells (Vero cells; ATCC CCL-81) or, in the case of ICP0 mutant viruses, human female osteosarcoma cells (U2OS cells; ATCC HTB-96). 293FT cells, a variant of the human kidney cell line 293, expressing simian virus 40 (SV40) large T antigen (provided by the Chen Luxenburg lab), were used for obtaining the VP16-expressing lentivirus. All cells were grown with Dulbecco’s modified Eagle medium (DMEM ×1; Gibco), supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin and streptomycin (10,000 units/mL and 10 mg/mL, respectively; Biological Industries, Israel).

The following materials were used during cell culture: the Ca salt (monohydrate) of HMB
(Ca-HMB) was purchased from Metabolic Technologies; Dulbecco’s Modified Eagle Medium
(DMEM) (1×) with glutamax, fetal bovine serum (FBS), horse serum (HS) and antibiotics
(AB) – penicillin–streptomycin and fungizone – were purchased from Gibco,
Life Technologies; penicillium crystalicum (AB) was purchased from Polfa Tarchomin; PBS,
protease from Streptomyces griseus and DMSO were purchased from Sigma
Aldrich. Tissue culture flasks Primaria (25, 75 cm²) and Collagen I Cellware
six-well plates were purchased from Becton Dickinson. Ca-HMB was transformed to the acid
form by acidification with 1 N-HCl. HMB was then extracted four times with diethyl
ether. The pooled organic layer was dried under vacuum for 24 h at 38 °C. The resulting
free acid was 99 % HMB as assessed by HPLC.