Fig. 8
The species and amounts of DVGs are altered under different infection environments and selection pressures. (A) Diagram depicting the primer sets used for determining the synthesis of BCoV DVGs in Figures (B)-(D). (B) Detection of DVGs synthesized in different cells infected with BCoV by RT-PCR. HRT-18, BHK, ML and A549 cells were infected with BCoV at an MOI of 0.1, followed by total cellular RNA collection at 24 hpi and RT-PCR. The “mock” indicates cells without infection with virus. (C) Detection of DVGs synthesized in different cells infected with BCoV-p95 by RT-PCR. HRT-18, BHK, ML and A549 cells were infected with BCoV-p95 at an MOI of 0.1, followed by total cellular RNA collection at 24 hpi and RT-PCR. (D) Detection of BCoV DVGs synthesized in HRT-18 cells infected with BCoV (0.1 MOI) and treated with antiviral remdesivir at final concentrations of 125, 250, 500 or 1000 nM. Total cellular RNA was collected at 48 hpi and DVGs were detected by by RT-PCR. (E) Diagram depicting the primer sets used for determining the MHV-A59 DVGs in Figure (F). (F) Detection of MHV-A59 DVGs synthesized in ML cells infected with MHV-A59 and treated with IFN β by RT-PCR. ML cells in 2 ml of DMEM were treated with IFN β at final concentrations of 103, 104 or 105 U/ml. After 16 h of treatment, IFN β-treated ML cells were infected with 0.1 MOI of MHV-A59 followed by total cellular RNA collection at 16 hpi. bp, base-pair; M, DNA size marker; m, mock-infected cells; HRT, human rectal tumor cells-18; BHK, baby hamster kidney cells; ML, mouse L cells; A549, adenocarcinomic human alveolar basal epithelial cells; hpi, hours postinfection; sgm, sgmRNA N; gm, genome; 18 S, 18 S rRNA