Ak 0529

HEK293T, Huh7.5.1, Vero, and HeLa cells were maintained in Dulbecco’s Modified Eagle Medium Hight Glucose (Gibco, Waltham, MA, USA) supplemented with 100 μg/mL streptomycin, 100 units/mL penicillin (Gibco, Waltham, MA, USA), and 10% fetal bovine serum (FBS) (ExCell Bio, Shanghai, China). The 293T-hACE2 cell line stably expressing human angiotensin-converting enzyme 2 (ACE2) was generated by lentiviral transduction of HEK293T cells with human ACE2, as previously described [38 (link),39 (link)]. HEp-2 cells were maintained in RPMI 1640 (Gibco, Waltham, MA, USA) supplemented with 100 μg/mL streptomycin, 100 units/mL penicillin, and 10% FBS. RSV entry inhibitors, TM353121, AK 0529, and RV521, were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Anti-VSV-G antibody [8G5F11] was purchased from Kerafast (Boston, MA, USA). The mouse anti-RSV-F serum and mouse anti-SARS-CoV-2 spike serum were prepared in our laboratory [39 (link)].

HEp-2 and HEK293T cells were purchased from the American Type Culture Collection (ATCC) and maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 2 mM l-glutamine. RSV A2 was obtained from Wuhan University, China. RSV A2-F_K394R was isolated and identified in our previous study (29 (link)). All the viruses were grown in HEp-2 cells and stored at −70°C. Viral titers were determined by plaque assay. Salvianolic acid R (LF-6) was isolated from the aerial part of Mesona chinensis Benth. (Lamiaceae) by the authors, following our previously reported procedure (30 (link)). The purity of LF-6 (>98%) was validated through high-performance liquid chromatography (HPLC) analysis. Ribavirin, heparin, and BMS-433771 were purchased from Sigma-Aldrich, Inc. TMC-353121, JNJ-53718678, GS-5806, and AK-0529 were purchased from MedChem Express, Inc.