Journal: PLOS Pathogens

Article Title: SARS-CoV-2 defective viral genomes from distinct genomic regions drive divergent interferon responses

doi: 10.1371/journal.ppat.1014099

Figure Lengend Snippet: (A) Genome schematics of DVG-B (left) and DVG-ΔN (right). (B) A549-ACE2 cells were infected with WT, DVG-B or DVG-ΔN at MOI 1 and cells were collected 24-hours post-infection for RT-qPCR. Expression of host genes and viral genes/eGFP were calculated relative to GAPDH. **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA with Tukey’s multiple comparisons test. N = 3, mean±SD. (C) A549-ACE2 cells were infected with WT (MOI 0.1), DVG-B or DVG-ΔN (MOI 1) and cells were collected 24-hours post-inoculation for confocal microscopy. Representative maximum intensity projections of merged dsRNA (red) staining. Nuclei were Hoechst stained (blue). GFP (green) indicates DVG-B and DVG-ΔN, respectively. Scale bar indicates 20μm. N = 4 fields for WT, DVG-B and DVG-ΔN, respectively. (D) A549-ACE2 cells were transduced with lentiviral vectors encoding SARS-CoV-2 orf9b (ORF9b) or an empty lentiviral vector control (EV). 48 hours post-transduction, cells were infected with DVG-ΔN (MOI 1). 24 hpi cells were collected for RT-qPCR. Expression of host and viral genes were calculated relative to GAPDH. *p < 0.05, by two-way ANOVA with Bonferroni’s multiple comparisons test. N = 3, mean±SD.

Article Snippet: Lentiviral constructs were obtained from addgene ( https://doi.org/10.1038/s41586-020-2286-9 ).

Techniques: Infection, Quantitative RT-PCR, Expressing, Confocal, Microscopy, Staining, Transduction, Plasmid Preparation, Control