hek-blue detection  (InvivoGen)

Journal: bioRxiv

Article Title: Dual roles for influenza A protein PA-X: limiting inflammatory response and disrupting MHC I antigen presentation in human respiratory epithelium

doi: 10.64898/2026.01.30.702929

Figure Lengend Snippet: A-B,D) ScRNAseq data from ALI cultures infected with Perth H3N2 WT or ΔX at MOI of 0.1, or mock infected as shown in were analyzed for changes in type I and III IFN production and responses (n = 2, from 2 separate donors). A) Heatmap showing average expression of type I and III IFNs at 1 and 3 DPI, separated in infected (influenza A virus-positive) and bystander (influenza A virus-negative) populations. Influenza A virus-positive cells were defined as cells of any subtype with at least 1% reads mapping to the influenza transcriptome. B) Heatmap showing average expression of type I and III IFNs at 1 DPI divided by cell type (assigned with ScType). C) ALI cultures were mock-infected or infected with Perth H3N2 WT or ΔX at MOI of 0.1. The basal medium was collected at the indicated timepoints. IFN-λ in the basal media was detected using HEK-Blue IFN-λ cells reporter cells. The IFN-λ concentration and the log2 fold-change in ΔX vs. WT infection are shown. (n=5-11 independent experiments, 5 donors, Donors 1-5). ns = p > 0.05, *= p ≤ 0.05, **= p ≤ 0.01, ***= p ≤ 0.001, two-way ANOVA with Šidák correction (IFN-λ concentration) or ratio T-test (Log2 fold-change). D) Heatmap showing average expression of ISGs at 1 and 3 DPI, separated in infected and bystander populations. ISG list was based on Schoggins et al. 2011.

Article Snippet: The following day, HEK-BlueTM IFN-λ Cell supernatant was incubated with QUANTI-BlueTM solution (InvivoGen) at 37°C in 5% CO 2 atmosphere for 30 minutes to 3 hours, then plates were read on the Cytation 5 plate reader (BioTek) at 620 and 655 nm.

Techniques: Infection, Expressing, Virus, Concentration Assay

Journal: bioRxiv

Article Title: Dual roles for influenza A protein PA-X: limiting inflammatory response and disrupting MHC I antigen presentation in human respiratory epithelium

doi: 10.64898/2026.01.30.702929

Figure Lengend Snippet: A-B,D) ScRNAseq data from ALI cultures infected with Perth H3N2 WT or ΔX at MOI of 0.1, or mock infected as shown in were analyzed for changes in type I and III IFN production and responses (n = 2, from 2 separate donors). A) Heatmap showing average expression of type I and III IFNs at 1 and 3 DPI, separated in infected (influenza A virus-positive) and bystander (influenza A virus-negative) populations. Influenza A virus-positive cells were defined as cells of any subtype with at least 1% reads mapping to the influenza transcriptome. B) Heatmap showing average expression of type I and III IFNs at 1 DPI divided by cell type (assigned with ScType). C) ALI cultures were mock-infected or infected with Perth H3N2 WT or ΔX at MOI of 0.1. The basal medium was collected at the indicated timepoints. IFN-λ in the basal media was detected using HEK-Blue IFN-λ cells reporter cells. The IFN-λ concentration and the log2 fold-change in ΔX vs. WT infection are shown. (n=5-11 independent experiments, 5 donors, Donors 1-5). ns = p > 0.05, *= p ≤ 0.05, **= p ≤ 0.01, ***= p ≤ 0.001, two-way ANOVA with Šidák correction (IFN-λ concentration) or ratio T-test (Log2 fold-change). D) Heatmap showing average expression of ISGs at 1 and 3 DPI, separated in infected and bystander populations. ISG list was based on Schoggins et al. 2011.

Article Snippet: HEK-BlueTM IFN-λ Cells (InvivoGen) were used to quantify type III interferon in media from the basal chamber of ALI culture wells.

Techniques: Infection, Expressing, Virus, Concentration Assay