Vero cell line

Vero cell line (ATCC, USA) was cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, USA) with 10% fetal bovine serum (FND500, ExCell Bio, Shanghai, China). The neuroblastoma cell line SK-N-SH (ATCC, HTB-11, American Type Culture Collection, Manassas, VA, USA) was propagated in Eagle’s minimal essential medium (MEM; GIBCO /Thermo Fisher Scientific, USA) supplemented with 10% FBS. In our previous studies, HSV-1 infection induced the biphasic F-actin dynamics in SK-N-SH cells [26 (link)]. In this study, the SK-N-SH cells were used to study the effect of amentoflavone on F-actin. HSV-1 strain F (ATCC, USA), initially obtained from Hong Kong University, was propagated in Vero cells and stored at −80 °C until use. HSV-1/Blue, a TK mutant derived from HSV-1 (KOS) and two acyclovir-resistant clinical HSV-1 strains HSV-1/106 and HSV-1/153 were kind gifts from Tao Peng, State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences. GFP-HSV-1, expressing a GFP-tagged viral protein Us11, was used to evaluate viral nuclear transport [27 (link)]. HSV-1 Us11 is a multifunctional late protein which can regulate the accumulation of RNA species and facilitate HSV-1 replication [28 (link)].

The African green monkey kidney Vero cell line (ATCC, Manassas, VA, USA), the human bronchial epithelial cell line 16HBE (ATCC), the human embryonic kidney cell line 293T (ATCC), the human glioma cell line Hs683 (ATCC), the human neuroblastoma cell line SH-SY5Y ATCC (and) the KMB17 cell line (IMB, CAMS, Yunnan, China) were maintained in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (HyClone, Logan, UT, USA). The culture medium was changed to DMEM supplemented with 2% fetal bovine serum after viral infection.

Vero cell line (ATCC, Manassas, VA, USA) was cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, Gaithersburg, MD, USA) with 10% fetal bovine serum (11011-8611, TIANHANG, Hangzhou, China). SH-SY5Y (ATCC, Manassas, VA, USA), a human neuroblastoma cell line was cultured in Dulbecco’s modified Eagle’s medium (DMEM; 8118305, GIBCO/Thermo Fisher Scientific, Gaithersburg, MD, USA) with 10% fetal bovine serum (FND500, ExCell Bio, Shanghai, China). HSV-1 strain F (ATCC, Manassas, VA, USA), initially obtained from Hong Kong University, was propagated in Vero cells and stored at −80 °C until use. EGFP-HSV-1, expressing an EGFP-tagged viral protein Us11, a kind gift from the College of Pharmacy, Jinan University (Guangzhou, China), was used to evaluate viral replication capacity [18 (link)].

Vero cell line (ATCC CCL-81), HCT-116 (ATCC CCL-247), A549 (ATCC CCL-185), MCF7 (ATCC HTB-22), H9c2 (ATCC CRL-1446), HepG2 (ATCC HB-8065), and Hacat (ATCC PCS-200-011) cells were purchased from American Type Culture Collection (ATCC, Manassas, Virginia, United States). Vero cells were grown in Eagle’s Minimal Essential Medium (EMEM) supplemented with 10% Fetal Bovine Serum (FBS), 100 mg/mL of streptomycin, 2 mM L-glutamine, and 100 IU/mL of penicillin-streptomycin solution in a humidified atmosphere with 5% CO₂ at 37 °C. HepG2 carcinoma cells were propagated in RPMI-1640 Medium containing 4.5 g/L glucose supplemented with 10% FBS, 100 IU/mL penicillin-streptomycin, and 2 mM L-glutamine. The other cell lines were propagated in Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% FBS, 2 mM L-glutamine and 100 IU/mL antibiotics. All the materials used for cell culture practice were purchased from Euroclone, Milan, Italy. HSV-1 (strain SC16), carrying a lacZ gene driven by the CMV IE-1 promoter to express β-galactosidase, was propagated on Vero cells monolayers. SARS-CoV-2 (clinical isolate, kindly provided by Lazzaro Spallanzani Hospital, Rome, Italy) was propagated on Vero cells as already reported [49 (link)]. All experimental work involving SARS-CoV-2 virus was performed in a biosafety level 3 (BSL3) containment laboratory.

Vero cell line (African green monkey kidney epithelial cells, ATCC CCL-81), HeLa cell line (ATCC, CCL-2), HEp-2 cell line (ATCC, CCL-23) and human neuroblastoma SK-N-SH cells (ATCC, HTB-11) were purchased from ATCC (Manassas, VA) or from Cell Bank of Chinese Academy of Sciences (Shanghai, China). Mouse embryo fibroblast (MEF) cells (isolated from C57BL/6J mouse embryos) were prepared in the lab following published protocol [64 (link)]. The cells were cultured in DMEM (high glucose) supplemented with 10% heat-inactivated fetal bovine serum (FBS), sodium pyruvate and non-essential amino acids (Life Technologies, Carlsbad, CA) in a humidified incubator at 37°C with 5% CO₂. HSV-1 strain F and HSV-2 strain G were propagated and titrated on Vero cells.