Journal: Antimicrobial Agents and Chemotherapy

Article Title: Selective Inhibition of Hepatitis C Virus Infection by Hydroxyzine and Benztropine

doi: 10.1128/AAC.02619-14

Figure Lengend Snippet: Selected compounds inhibit early aspects of HCV infection. (A) Huh7 cells were inoculated at a MOI of 10 in the presence of perphenazine (5 μM), hydroxyzine (5 μM), benztropine (7.5 μM), or DMSO as a control. Samples of cells and supernatants were collected to determine infectivity titers as well as intracellular HCV RNA levels by titration and RT-qPCR, respectively, at the indicated time points. (B) Inhibition of HCVtcp infection. Huh-7 cells were inoculated with HCVtcp in the presence of perphenazine (5 μM), hydroxyzine (5 μM), and benztropine (7.5 μM), and samples of the cells were assayed for luciferase activity 48 h postinoculation. Data are shown as averages and standard deviations of the results of two independent experiments performed in triplicate (n = 6). (C) Treatment of Huh-7 cells electroporated with a subgenomic HCV RNA replicon bearing a luciferase gene with perphenazine (5 μM), hydroxyzine (5 μM), benztropine (7.5 μM), or DMSO and 2mAde (10 μM) as negative and positive controls, respectively. Data are shown as averages and standard deviations of the results of two independent experiments performed in duplicate (n = 4). The statistical significance of the differences with the control data set was determined using Student's t test (*, P < 0.05; **, P < 0.01).

Article Snippet: 2mAde (2′-c-methyladenosine) was purchased from BOC Sciences (Shirley, NY).

Techniques: Infection, Control, Titration, Quantitative RT-PCR, Inhibition, Luciferase, Activity Assay

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Selective Inhibition of Hepatitis C Virus Infection by Hydroxyzine and Benztropine

doi: 10.1128/AAC.02619-14

Figure Lengend Snippet: Impact of the selected compounds on persistently infected cell cultures. Persistently infected cells were treated for 48 h with perphenazine (5 μM), hydroxyzine (5 μM), benztropine (7.5 μM), or DMSO and 2mAde (10 μM) as negative and positive controls, respectively. Samples of cells and supernatants were collected to determine infectivity titers as well as intracellular HCV RNA levels by titration and RT-qPCR, respectively. Viruses present in the supernatant were partially purified using microfiltration devices to eliminate residual compound present in the supernatant. Infectivity titers were determined in the purified material. Data are shown as averages and standard deviations of the results of two independent experiments performed in triplicate (n = 6). The statistical significance of the differences with the control data set was determined using Student's t test (*, P < 0.05; **, P < 0.01).

Article Snippet: 2mAde (2′-c-methyladenosine) was purchased from BOC Sciences (Shirley, NY).

Techniques: Infection, Titration, Quantitative RT-PCR, Purification, Control

Journal: bioRxiv

Article Title: The cellular poly(rC)-binding protein 2 prevents the early steps of hepatitis C virus virion assembly

doi: 10.1101/2022.04.12.488029

Figure Lengend Snippet: (A) Schematic representation of the experimental approach for 2’CMA experiments: two days post-siRNA transfection, Huh-7.5 cells were infected with JFH-1 T (MOI = 0.05). Three days post-infection, cells were treated with 2 ’ CMA or DMSO (control). Total RNA, intracellular and extracellular virus was collected at t = 0, 6 and 12 h post-treatment. (B) Quantitative RT-PCR analysis after 2’CMA treatment. (C) Intracellular viral titers and (D ) intracellular virus accumulation rate calculated by linear regression. (E) Extracellular viral titers and (F) virus secretion rates calculated by linear regression. (G) Model of PCBP2’s role in the HCV life cycle: PCBP2 normally inhibits the first stage of virion assembly, where the viral NS5A protein transfers the viral RNA to the core protein. All data are representative of three independent replicates and error bars in (B), (C) and (E) represent the standard deviation of the mean. Error bars in (D) and (F) represent the slopes of the linear regressions ± standard error. P-values were calculated by two-way ANOVA (ns, not significant; ** p < 0.01; **** p < 0.0001).

Article Snippet: Three days post-infection, the media on these cells was changed for complete Huh-7.5 media with 5 μM 2’CMA (2’C-methyladenosine, Carbosynth), an HCV NS5B polymerase inhibitor, or DMSO (vehicle control) ( ).

Techniques: Transfection, Infection, Control, Virus, Quantitative RT-PCR, Standard Deviation

Journal: iScience

Article Title: Experimental verification of strain-dependent relationship between mycovirus and its fungal host

doi: 10.1016/j.isci.2023.107337

Figure Lengend Snippet:

Article Snippet: 7-deaza-2′-C-methyladenosine , Biosynth Ltd. , ND08351.

Techniques: Recombinant, Multiplex Assay, Isolation, Virus, Sequencing, Software