recombinant human ifn γ  (MedChemExpress)

Journal: Advanced Science

Article Title: EGR Proteins Mediate Interferon‐Independent Anti‐HSV‐1 Responses Through Viral and Host Targets

doi: 10.1002/advs.202515546

Figure Lengend Snippet: EGR1 suppresses HSV‐1 replication by enhancing IRF7 expression. (A) Neuro‐2a cells were transfected with the indicated siRNAs for 24 h, then infected with HSV‐1 (MOI = 0.2) for 48 h before virus titration. (B) Left, Neuro‐2a cells were transfected with the indicated siRNAs for 18 h, then transfected with the indicated plasmids for 20 h, then infected with HSV‐1 (MOI = 0.2) for 44 h before virus titration. Right, Neuro‐2a cells were transfected with the indicated siRNAs for 16 h, then transfected with the indicated plasmids for 22 h, then infected with HSV‐1 (MOI = 0.5) for 24 h before virus titration. (C) Left, Neuro‐2a cells were transfected with 400 ng/mL of indicated plasmids expressing flag‐tagged proteins for 24 h before Western blot analysis using a flag antibody. Right, Neuro‐2a cells were transfected with the indicated plasmids for 24 h, then infected with HSV‐1 (MOI = 0.2) for 48 h before virus titration. (D) RT‐qPCR analysis of IRF7 mRNA at the indicted times after HSV‐1 infection of Neuro‐2a cells (MOI = 10). (E) N2A‐EGR1‐KO1 cells were co‐transfected with 300 ng/mL of pcDNA or pIRF7, 100 ng/mL of the ISRE‐luc plasmid and 50 ng/mL of the RL‐CMV plasmid for 24 h before luciferase assays. (F) N2A‐EGR1‐KO1 cells were untreated or pretreated with IFN‐α (500 IU/mL) for 12 h, then co‐transfected with 300 ng/mL of pcDNA or pEGR1, 100 ng/mL of ISRE‐luc, 50 ng/mL of RL‐CMV in the absence (left) or presence (right) of IFN‐α for 24 h before luciferase assays. (G) Neuro‐2a cells were transfected with the indicated plasmids for 22 h, then treated with DMSO or 2 µM IFNAR1 inhibitor (IFN alpha‐IFNAR‐IN‐1) for 16 h, and then infected with HSV‐1 (MOI = 0.2) in the presence of DMSO or the inhibitor for 40 h before virus titration. n = 3 biologically independent samples for all panels. Data were analyzed by one‐way ANOVA with Dunnett's multiple comparisons tests (A, C, D) or two‐way ANOVA with Sidak's multiple comparisons tests (B, G), two‐tailed unpaired t tests (E, F) and are presented as mean ± SD.

Article Snippet: The following inhibitors were used at the following final concentrations: PDK1 inhibitor (BX‐795, 1 μ m , MedChemExpress), PI3K inhibitor (PI‐103, 500 n m , MedChemExpress), MEK1/2 inhibitor (Binimetinib, 1 μ m , MedChemExpress), AKT inhibitor (MK‐2206, 1.25 μ m , MedChemExpress), ATM inhibitor (KU‐55933, 20 μ m , MedChemExpress), FNAR1 inhibitor (IFN alpha‐IFNAR‐IN‐1, HY‐12836A, 2 μ m , MedChemExpress), and JNK inhibitor (SP600125, 10 μ m , MedChemExpress).

Techniques: Expressing, Transfection, Infection, Virus, Titration, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Luciferase, Two Tailed Test

Journal: Immunity, Inflammation and Disease

Article Title: Activated Peripheral CD8 + T Lymphocytes Inhibit the Proliferation of Hippocampal Neural Stem Cells via the IFN‐γ/JAK/STAT Signaling Pathway

doi: 10.1002/iid3.70287

Figure Lengend Snippet: Activated T cells inhibit NSC proliferation by releasing IFN‐γ. (A) IFN‐γ secretion concentration at different scales ( n = 3); (B) Immunofluorescence showing the presence of IFNGR on NSCs under different treatment conditions, scale bar = 100 μm; (C and E) Representative immunofluorescence images of EdU+ NSCs under different treatment conditions. The image at the bottom left showed a magnification of the point of the arrow. Scale bar = 100 μm; (D and F) Statistical diagram of EdU+ NSCs under different treatment conditions ( n = 5). UAT, unactivated CD8 + T lymphocytes; AT, activated CD8 + T lymphocytes; AT + N, activated CD8 + T lymphocytes and IFN‐γ neutralizing antibodies. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: IFN‐γ (20 pg/mL, Servicebio, China) or AG490 (a JAK inhibitor, 10 μmol/L, MCE, USA) was added into the culture medium if necessary.

Techniques: Concentration Assay, Immunofluorescence

Journal: Immunity, Inflammation and Disease

Article Title: Activated Peripheral CD8 + T Lymphocytes Inhibit the Proliferation of Hippocampal Neural Stem Cells via the IFN‐γ/JAK/STAT Signaling Pathway

doi: 10.1002/iid3.70287

Figure Lengend Snippet: IFN‐γ secreted by activated CD8 + T lymphocytes causes the phosphorylation of JAK2 and STAT1 in NSCs. (A) Representative western blot image of p‐Jak1 and p‐Jak2 expression in NSCs under different treatment conditions; (B and C) Relative expression of p‐Jak1 and p‐Jak2 proteins; (D) Representative western blot image of p‐STAT1‐tyr, p‐STAT1‐ser and t‐STAT1 expression in NSCs under different treatment conditions; (E–G) Relative expression of p‐STAT1‐tyr, p‐STAT1‐ser and t‐STAT1 proteins; (H) Representative western blot image of p‐STAT3‐tyr, p‐STAT3‐ser and t‐STAT3 expression in NSCs under different treatment conditions; (I–K) Relative expression of p‐STAT1‐tyr, p‐STAT1‐ser and t‐STAT1 proteins. AT, activated CD8 + T lymphocytes; AT + N, activated CD8 + T lymphocytes and IFN‐γ neutralizing antibodies; UAT, unactivated CD8 + T lymphocytes. N = 3/group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: IFN‐γ (20 pg/mL, Servicebio, China) or AG490 (a JAK inhibitor, 10 μmol/L, MCE, USA) was added into the culture medium if necessary.

Techniques: Phospho-proteomics, Western Blot, Expressing