Journal: Frontiers in Immunology

Article Title: Identification of interferon-stimulated genes with modulated expression during hepatitis E virus infection in pig liver tissues and human HepaRG cells

doi: 10.3389/fimmu.2023.1291186

Figure Lengend Snippet: List of primers used for the quantification of human gene expression by RT-qPCR.

Article Snippet: The membrane was then incubated with the required dilution of specific antibodies raised against ORF2 (mouse, 1/2000 dilution, clone 1E6, Merck Millipore), actin (mouse, 1/2000 dilution, clone M2, Sigma), DDX58 (mouse, 1/1000 dilution, clone Alme-1, AdipoGen Life Sciences), IFIH1 (rabbit, 1/1000 dilution, AT113, ALX-210-935, Enzo Life Sciences), or IRF1 (rabbit, 1/1000 dilution, VPA00801, Bio-Rad).

Techniques: Gene Expression

Journal: Frontiers in Immunology

Article Title: Identification of interferon-stimulated genes with modulated expression during hepatitis E virus infection in pig liver tissues and human HepaRG cells

doi: 10.3389/fimmu.2023.1291186

Figure Lengend Snippet: Validation of the PCR array data. (A) HepaRG cells were infected or not with HEV-3f for 100 days at MOI 100 and the expression of selected genes shown by PCR array to be up-regulated or unchanged was analyzed by RT-qPCR. The results shown are the geometric means ± SD from 3 to 4 replicate samples and are representative of 2 independent experiments. GUSB , B2M and GAPDH were used as reference genes. Unpaired t-test with Welch’s correction Mann-Whitney test, *: p<0.05 (B) Immunoblot showing the expression of the protein encoded by IFIH1 , DDX58 and IRF1 in HepaRG cells infected with HEV-3f or not (NI) for 100 days at MOI 100 for 3 replicate samples. Representative blots from 2 independent experiments are shown. ORF2 and actin protein levels were also detected as control of infection and loading, respectively. (C) Detection of CXCL10 in the supernatant of non-infected HepaRG (NI) or HepaRG cells infected with HEV-3f for 100 days at MOI 100 (GE/cell) by ELISA. The results shown are the means ± SD from 4 replicate samples and are representative of 2 independent experiments. Unpaired t-test with Welch’s correction, *: p<0.05.

Article Snippet: The membrane was then incubated with the required dilution of specific antibodies raised against ORF2 (mouse, 1/2000 dilution, clone 1E6, Merck Millipore), actin (mouse, 1/2000 dilution, clone M2, Sigma), DDX58 (mouse, 1/1000 dilution, clone Alme-1, AdipoGen Life Sciences), IFIH1 (rabbit, 1/1000 dilution, AT113, ALX-210-935, Enzo Life Sciences), or IRF1 (rabbit, 1/1000 dilution, VPA00801, Bio-Rad).

Techniques: Biomarker Discovery, Infection, Expressing, Quantitative RT-PCR, MANN-WHITNEY, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Virology

Article Title: Differential Responses by Human Respiratory Epithelial Cell Lines to Respiratory Syncytial Virus Reflect Distinct Patterns of Infection Control

doi: 10.1128/JVI.02202-17

Figure Lengend Snippet: Summary of differential gene expression kinetics between A549 and BEAS-2B cells in response to low doses of RSV

Article Snippet: Levels of IRF1, IRF9, and RIG-I were visualized by chemiluminescence using rabbit anti-IRF1 monoclonal antibody (MAb) (clone D5E4; Cell Signaling Technology), rabbit anti-ISGF-3γ polyclonal antibody (PAb) (Santa Cruz, Dallas, TX), mouse anti-RIG-I MAb (clone 1c3; Millipore), and mouse anti-β-actin MAb (clone C4; Santa Cruz) as a control for protein loading.

Techniques: Expressing

Journal: Journal of Virology

Article Title: Differential Responses by Human Respiratory Epithelial Cell Lines to Respiratory Syncytial Virus Reflect Distinct Patterns of Infection Control

doi: 10.1128/JVI.02202-17

Figure Lengend Snippet: RSV-infected BEAS-2B cells express more IRF-1 but activate less IRF-3 and NF-κB than A549 cells. A549 or BEAS-2B cells were mock infected (medium only) or infected with rgRSV at an MOI of 0.1 or 0.3 for the indicated times. (A) Transcription factor gene expression was measured by RT-qPCR. The data shown are means ± SD from the results of four experiments performed in duplicate. Statistical differences between cell lines at each time point were calculated using a one-way ANOVA model without adjustment for mock infection on log10-transformed values as described in Materials and Methods. *, P < 0.05 for A549 compared to BEAS-2B mock infection at 0 h; ++, P < 0.05 for A549 compared to BEAS-2B at MOIs of 0.1 and 0.3; +, P < 0.05 at an MOI of either 0.1 or 0.3. (B and D) IRF3 dimerization (B) and IRF1 protein expression (D) were measured by Western blotting. The data represent the results of experiments on two separate cultures. (C) EMSA was performed to determine NF-κB activation.

Article Snippet: Levels of IRF1, IRF9, and RIG-I were visualized by chemiluminescence using rabbit anti-IRF1 monoclonal antibody (MAb) (clone D5E4; Cell Signaling Technology), rabbit anti-ISGF-3γ polyclonal antibody (PAb) (Santa Cruz, Dallas, TX), mouse anti-RIG-I MAb (clone 1c3; Millipore), and mouse anti-β-actin MAb (clone C4; Santa Cruz) as a control for protein loading.

Techniques: Infection, Expressing, Quantitative RT-PCR, Transformation Assay, Western Blot, Activation Assay