RNA-seq analysis and validation of simeprevir-mediated
host response
and antiviral activity. (A) Schematic representation of RNA-seq sample
preparation. Treatment sequence and incubation time of simeprevir
and SARS-CoV-2 was indicated with arrows and legends. (B) Venn diagrams
showing differentially expressed genes (DEGs) comparing simeprevir-treated
(1.1 or 3.3 μM), infected, and mock-infected samples. (C) Bubble
plot of top 20 hits of positively enriched reactome gene sets under
simeprevir treatment using gene set analysis (GSEA). Enriched gene
sets were filtered with criteria false discovery rate (FDR) q-value < 0.25 and nominal p-value <
0.05 before ranked with their normalized enrichment scores (NES).
(D) Enrichment plots of GSEA results using gene ontology (GO) gene
sets. (E) Clustered heatmap showing the row-normalized expression
level of genes belonging to GO term “response to type I interferon”.
(F) Viral titration assay using A549-ACE2 cells treated with 4 μM
simeprevir and/or 1 μM JAK inhibitor I. (G) Relative RNA level
of ISG15 in uninfected or infected A549-ACE2 cells, treated with 4
μM simeprevir and/or 1 μM JAK inhibitor I. Data points
in all plots represent mean ± S.D. For all data points, n = 3 replicates; * p-value < 0.05,
*** p-value < 0.005.