Journal: Virologica Sinica

Article Title: Two orthobunyaviruses: OYAV and EBIV co-opt the AhR-CYP1A1 axis to suppress type I interferon responses

doi: 10.1016/j.virs.2026.04.001

Figure Lengend Snippet: CYP1A1 enhances OYAV and EBIV infection. A, B Viral RNA levels and progeny viral titers were measured in CYP1A1-overexpressing HeLa cells, infected with OYAV (MOI = 0.01, 24 hpi) and EBIV (MOI = 0.01, 24 hpi), via RT-qPCR and plaque formation assay. C, D CYP1A1 knockout (KO) HepG2 clonal lines generated by CRISPR-Cas9 were infected with OYAV (MOI = 0.1, 24 hpi) and EBIV (MOI = 0.1, 24 hpi). Viral RNA levels and progeny viral titers were measured via RT-qPCR and plaque formation assay. E, F Viral RNA levels and progeny viral titers were measured in CYP1A1-konckout-complemented and WT HepG2 cells, infected with OYAV (MOI = 0.1, 24 hpi) and EBIV (MOI = 0.1, 24 hpi), via RT-qPCR and plaque formation assay. G Growth rate of CYP1A1 knockout (KO) HepG2 clonal cell lines was determined by plotting growth curves at days 1–6 of cell growth. H Progeny viral titers were measured in HepG2 cells, treated or untreated with 1 μM Ethinylestradiol (EE) or 10 μM Alpha-Naphthoflavone (ANF) and infected with OYAV and EBIV (MOI = 0.1, 24 hpi), via plaque formation assay. The threshold cycle (Ct) values of the target genes were normalized to that of the housekeeping gene GAPDH and presented as relative RNA levels, with the normalized level of the target gene in the WT and Vector group set as 1. Data represent mean ± SD (n = 3; two-tailed t -test for A, B ; one-way ANOVA test for C, D, E, F, H ).

Article Snippet: Ethinylestradiol; Alpha-Naphthoflavone and CH223191 were purchased from MedChemExpress.

Techniques: Infection, Quantitative RT-PCR, Plaque Formation Assay, Knock-Out, Generated, CRISPR, Plasmid Preparation, Two Tailed Test

Journal: Virologica Sinica

Article Title: Two orthobunyaviruses: OYAV and EBIV co-opt the AhR-CYP1A1 axis to suppress type I interferon responses

doi: 10.1016/j.virs.2026.04.001

Figure Lengend Snippet: AhR-CYP1A1 axis suppress type I interferon responses to regulate OYAV and EBIV infections. A, B Detect the activation status of proteins (STAT1, TBK1) related to the type I interferon pathway in wild-type and CYP1A1 knockout HepG2 cells with EBIV (MOI = 1) infection after 0, 12, 24 h. The gray values were measured. C Heatmap visualization of differentially expressed genes. The heatmap displays the expression patterns of the top 11 differentially expressed, dirlenectly interferon-related genes in wild-type (WT) and CYP1A1 knockout (KO) HepG2 cells under EBIV infection conditions (MOI = 1, 24 hpi). Colors represent the expression level of each gene (row) processed by log 10 (FPKM). Shown are genes with |Log 2 FC| > 1 and adjusted P -value <0.05. D Viral replication, IFIT3, IFIT5, IFI16, IFI44, IFITM1, OASL, OAS2, RIGI, STAT1, and STAT2 mRNA levels in wild-type (WT) and CYP1A1 knockout (KO) HepG2 cells with EBIV infection (MOI = 1, 24 hpi). E Viral RNA levels were measured in CYP1A1-overexpressing Vero cells, infected with OYAV (MOI = 0.01, 24 hpi) and EBIV (MOI = 0.01, 24 hpi). F Viral RNA levels were measured in Vero cells, treated or untreated with 1 μM Ethinylestradiol or 10 μM Alpha-Naphthoflavone and infected with OYAV and EBIV (MOI = 0.01, 24 hpi). G Proposed model: Viral infection activates AhR nuclear translocation, upregulating CYP1A1 to inhibit IFN-I production. The threshold cycle (Ct) values of the target genes were normalized to that of the housekeeping gene GAPDH and presented as relative RNA levels, with the normalized level of the target gene in the Vector, DMSO and WT group set as 1. Data represent mean ± SD (n = 3; two-tailed t -test for B, D, E , one-way ANOVA test for F ).

Article Snippet: Ethinylestradiol; Alpha-Naphthoflavone and CH223191 were purchased from MedChemExpress.

Techniques: Activation Assay, Knock-Out, Infection, Expressing, Translocation Assay, Plasmid Preparation, Two Tailed Test

Journal: Virologica Sinica

Article Title: Two orthobunyaviruses: OYAV and EBIV co-opt the AhR-CYP1A1 axis to suppress type I interferon responses

doi: 10.1016/j.virs.2026.04.001

Figure Lengend Snippet: CYP1A1 exhibits broad spectrum proviral activity. A Viral RNA levels were measured in CYP1A1-konckout HepG2 cells with RVFV, SFTSV, JEV, ZIKV, or TMUV infection (MOI = 0.1, 24 hpi). B Viral RNA levels were measured in HepG2 cells, treated or untreated with 1 μM Ethinylestradiol (EE) and infected with RVFV, SFTSV, JEV, ZIKV, or TMUV infection (MOI = 0.1, 24 hpi). C, D Detected AhR and CYP1A1 expression levels of HepG2 stimulated by RVFV, SFTSV, JEV, ZIKV and TMUV (MOI = 1) infection after 24 h. The threshold cycle (Ct) values of the target genes were normalized to that of the housekeeping gene GAPDH and presented as relative RNA levels, with the normalized level of the target gene in the WT, Mock and DMSO group set as 1. Data represent mean ± SD (n = 3 independent experiments; one-way ANOVA test for A, B, C, D ).

Article Snippet: Ethinylestradiol; Alpha-Naphthoflavone and CH223191 were purchased from MedChemExpress.

Techniques: Activity Assay, Infection, Expressing