Figure 4. Detection of interferon-stimulated circRNAs (ISCs).

A-E) MRC-5, LEC, or Akata- cells were treated with recombinant interferons for 48 hours (n=3). MRC-5 and LEC were treated with IFN-β and -γ (25 ng/mL conc). Akata- were treated with IFN-β (10 ng/mL). Bulk RNA-Seq was performed and data was normalized relative to ERCC spike-in controls. A, C) Volcano plots for ERCC normalized mRNA or circRNA reads. P-values were calculated using EdgeR. Log₂FC was calculated relative to a paired untreated sample. B, D) Venn diagrams of significantly upregulated circRNA or mRNAs. DECs had a raw BSJ count across the sample set >10, Log2FC>0.5 and EdgeR p-value <0.05. DEGs had a Log₂FC>0.5 and EdgeR p-value <0.05. E) Log₂FC (stimulated/untreated) was plotted for DECs in B and D relative to their colinear gene reads. R² values are from linear regression analysis. F) Overlap of circRNAs upregulated during herpesvirus infection (Fig. 1) or interferon-stimulation (Fig 4B, D).