wildtype hadv c5  (ATCC)

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: (A): A549 clone 2E10 was treated with IFN⍺/IFNγ for 16h then infected as indicated. Infection was assessed 72 hpi by flow cytometry. (B): RNA-seq analysis heatmap depicting normalized expression of selected ISGs across four A549 SCCs (1B7, 1C10, 2E10, 3G7) after 16h mock/IFNα/IFNγ treatment. TPM; transcripts per million. (C): Venn diagram of overlap between differentially expressed genes (>4-fold change and padj < 0.05) following IFNα/IFNγ treatment. (D): Schematic of screen setup. (E): Screen results, where the x axis corresponds to each unique gene in the library, and the y axis denotes the relative fold change. Differentially represented genes in infected versus uninfected populations (p<0.01) are labeled in magenta, non-targeting controls are labeled in blue. (F): Flow cytometry of knockout cells treated with IFNα or IFNγ for 16h and infected with HCoV-OC43 for 72h (normalized to EV control).

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Infection, Flow Cytometry, RNA Sequencing, Expressing, Labeling, Knock-Out, Control

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: (A): A549 single cell clones (n=20) treated with IFN⍺/IFNγ as indicated for 16h, then infected with rVSV/NanoLuc (MOI 0.003). At 24 hpi cells were lysed and luciferase activity measured. (B): RNA-seq analysis of four A549 SCCs (1B7, 1C10, 2E10, 3G7) treated with IFN for 16h. Volcano plots show gene expression changes induced by IFNα or IFNγ compared to mock-treated controls. (C): Flow cytometry at 72 hpi of A549 cells (clone 2E10) treated with IFN⍺/IFNγ as indicated for 16 hours, and infected with the indicated virus to produce ∼1% infection after treatment under conditions used for the screen. (D): Genomic DNA was extracted from screened cells. DNA concentration was measured, and qPCR performed with the indicated primers to assess the quality of the library. (E): Infected A549 cells (clone 2E10) at 3 dpi were fixed for the indicated number of days (dpf: days post fixation), and percent infection was determined by flow cytometry. (F): Results of targeted ISG screens against four viruses, analyzed using PinAPL-py. Relative fold change represents the average fold change of guides targeting a gene in the infected population, minus the mean fold change of the control, divided by the standard deviation of the control (sigmaFC). (G): Selected ISGs (19) were individually knocked out in A549 cells. Cells were treated with IFNα or IFNγ for 16h then infected with the indicated virus for 72h. Infection levels were measured by flow cytometry and normalized to the empty vector (EV) control.

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Single Cell, Clone Assay, Infection, Luciferase, Activity Assay, RNA Sequencing, Gene Expression, Flow Cytometry, Virus, Concentration Assay, Control, Standard Deviation, Plasmid Preparation

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: (A): RNF213 KO (n=20) or EV (n=7) A549 SCCs were treated as indicated for 16h, infected with HCoV-OC43 at MOI 0.1 for 72h and percent infected cells determined by flow cytometry. Each dot represents an independent SCC (representative experiment, n=3; statistics used each clone as an data point, +/- SD, p values shown: student’s t-test). (B) : Growth curve of HCoV-OC43 in A549 EV or RNF213 KO cells pretreated, or not, for 16h with IFNγ and infected at MOI 0.1 (n=3). (C): Cells were pretreated with IFNγ for 16h and infected with HCoV-OC43 at MOI 5 for 24h. Labeled extracellular virions were run on an SDS-PAGE gel (representative image, n=3). (D): A549 cells were transfected with siRNAs against RNF213 or a non-targeting control (NT), pretreated with IFNγ for 16h, and plaque assay performed for rHCoV-OC43/eGFP (representative image, n=3, scale 1 mm). (E): Quantification of plaque size (representative experiment shown +/- SD, p values shown: 2-way ANOVA). (F): Quantification of plaque number (n=3, +/- SD, p values shown: 2-way ANOVA). (G): siRNA transfected A549 cells were pretreated or not with IFNγ and infected with HCoV-OC43. Percentage infected cells at 72 hpi was determined by flow cytometry (n=3 +/-SD), (H): Same as (G) , in Vero cells (I): Schematic of human RNF213 and locations of CRISPR-mediated editing. (J): Flow cytometry of A549 SCCs containing the indicated RNF213 mutations, pretreated with IFNγ (16h) and infected with HCoV-OC43 at MOI 0.1 for 72h. Each dot represents an independent SCC (representative experiment, n=3; statistics based on each clone as an independent experiment, +/- SD, p values shown: 2-way ANOVA). EV: empty vector. KO: knockout. MOI: multiplicity of infection. PFU: plaque forming units. Tri: Trimer. Mono: Monomer. SCC: single-cell clone.

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Infection, Flow Cytometry, Labeling, SDS Page, Transfection, Control, Plaque Assay, CRISPR, Plasmid Preparation, Knock-Out, Single Cell

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: (A): Quantification of fluorescent signal for each of the indicated bands from (n=3, +/- SD, p-values displayed: student’s t-test). (B): Calculation of the ratio of each other protein to M (n=3, +/- SD, p-values displayed: student’s t-test). (C): Flow cytometry analysis of A549 RNF213 knock-out cells containing knock-in mutations in the guide RNA target sequence. Cells were pretreated for 16h with IFNγ and infected with HCoV-OC43 at MOI 0.1 for 72 hours (n=3, +/- SD, p-values shown: 2-way ANOVA). (D): Western blot of EV or RNF213 KO A549 cells pretreated with IFN for 24 hours (representative images shown, n=3). (E): Western blot of the indicated cells transfected with siRNAs against RNF213 or a non-targeting control, then pretreated with IFNγ for an additional 24 hours (representative images shown, n=3). (F-G): Flow cytometry of the indicated cells transfected with siRNAs, pretreated with IFNγ for 16 hours, and infected with HCoV-OC43 (F) or the indicated virus (G) at the indicated MOI for 72h, except VSV which was infected for 24h (n=3. +/- SD). (H): Western blot of A549 cells with the indicated point mutations pretreated with IFNγ for 24 hours (representative images shown, n=3).

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Flow Cytometry, Knock-Out, Knock-In, Sequencing, Infection, Western Blot, Transfection, Control, Virus

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: (A): Schematics of recombinant rHCoV-OC43 viruses expressing reporter proteins. (B): Flow cytometry analysis at 24 hpi of EV or RNF213 KO A549 SCC cells pretreated with IFNγ for 16h, and infected with rHCoV-OC43/eGFP at MOI 0.1 in the presence or absence of 15% normal human serum added at 4 hpi (NHS) (n=3, +/- SD, p-values displayed, student’s t-test). (C): Same as B, except that cells were infected at MOI 1, 15% NHS added at 4 hpi, and qPCR was performed for genomic and subgenomic RNA (n=3, +/- SD, p-values displayed: student’s t-test). (D): Schematics of entry (left) and primary translation (right) assays. (E-H): Vero cells were transfected with the indicated siRNAs, treated with IFNγ or no IFN for 16 hours, then infected. (E): Infection of cells expressing LgBiT with rHCoV-OC43/M-HiBiT at MOI 0.04. Percent infection at 72 hpi determined by flow cytometry (n=3, +/- SD, p-values displayed: 2-way ANOVA). (F): Infection of cells expressing LgBiT with rHCoV-OC43/M-HiBiT at MOI 3. Luminescence was measured at the indicated times post infection (n=3, +/- SD). (G): Infection with rHCoV-OC43/NLuc-NSP3/eGFP at MOI 0.56. Percent infection at 48 hpi determined by flow cytometry (n=3, +/- SD, p-values displayed: 2-way ANOVA). (H): Infection with rHCoV-OC43/Nluc-NSP3/eGFP at MOI 1. Luminescence was detected at the indicated times post infection (n=3, +/-SD).

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Recombinant, Expressing, Flow Cytometry, Infection, Transfection

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: (A): Flow cytometry analysis of A549 cells transfected with siRNAs, treated with IFNγ for 16 hours, and infected with the indicated virus at the indicated MOI for 72h (n=3, +/- SD). (B): Same in Vero cells, except rHCoV-OC43/NLuc-NSP3/eGFP was infected for 48h. (C): Flow cytometry analysis of 293T cells infected with rHCoV-OC43/eGFP at MOI 0.1 for 72h, with normal human serum added at 4 hpi (representative experiment, n=3). (D): Same with A549 cells. (E): Flow cytometry analysis of Vero cells infected with rHCoV-OC43/eGFP at MOI 1 for 24h with soluble heparin added concurrently with infection. (F): Vero cells expressing LgBiT were infected with rHCoV-OC43/M-HiBiT at MOI 1 for 24h in the presence of the luciferase detection reagent endurazine. Soluble heparin was added at 0 hpi and/or at 24 hpi, and luminescence detected at 25 hpi (n=3. +/-SD). (G): qPCR of A549 cells infected with HCoV-OC43 at MOI 0.1 for 24h in the presence of Molnupiravir (representative images, n=3). (H): Western blot of EV or RNF213 KO A549 cells infected for the indicated time with HcoV-OC43 and 200 μM Molnupiravir (representative images, n=3.

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Flow Cytometry, Transfection, Infection, Virus, Expressing, Luciferase, Western Blot

Journal: bioRxiv

Article Title: Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

doi: 10.64898/2026.06.02.729663

Figure Lengend Snippet: A549 cells were treated with IFNγ for 24 hours and stained for the indicated targets (red, green), and Hoeschst 33342 (blue) for nuclei. (A): EV or RNF213 KO cells, stained for endogenous RNF213 (green). Two leftmost images are maximum intensity projection of a z-stack, right image is a selection from a 5x5 mosaic image of a single focal plane. (B): Cells cultured with Lipi-Deep Red to stain lipid droplets (red) and stained for endogenous RNF213 (green). (C): Cells containing the indicated knock-in mutation in RNF213 stained with an antibody against endogenous RNF213 (green). Except as noted, maximum intensity projection of a z-stack is shown (representative images, n=3; scale bars are 10 µm for large images and 3 µm for magnified insets).

Article Snippet: Adenovirus 5 was purchased from ATCC (VR-1516) and propagated in A549 cells.

Techniques: Staining, Selection, Cell Culture, Knock-In, Mutagenesis

Journal: GMS Hygiene and Infection Control

Article Title: Virucidal efficacy of a UV-C disinfection system for semi-critical medical devices according to EN 14476

doi: 10.3205/dgkh000622

Figure Lengend Snippet: lg TCID 50 /mL titers of Adenovirus Type 5 on glass test surface at initial inoculation, post-drying, and after UV-C treatment

Article Snippet: The following indicator viruses were tested in accordance with EN 14476 requirements: Adenovirus Type 5 (ATCC VR-5), Poliovirus Type 1 LSc-2ab (RVB-1260), Murine Norovirus strain S99 (RVB-651), and Simian Virus 777 (SV40).

Techniques:

Journal: GMS Hygiene and Infection Control

Article Title: Virucidal efficacy of a UV-C disinfection system for semi-critical medical devices according to EN 14476

doi: 10.3205/dgkh000622

Figure Lengend Snippet: lg TCID 50 /mL titers of Adenovirus Type 5 on TPE-test surface at initial inoculation, post-drying, and after UV-C treatment

Article Snippet: The following indicator viruses were tested in accordance with EN 14476 requirements: Adenovirus Type 5 (ATCC VR-5), Poliovirus Type 1 LSc-2ab (RVB-1260), Murine Norovirus strain S99 (RVB-651), and Simian Virus 777 (SV40).

Techniques: