Mouse IFN-gamma Antibody (H22)

IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Mouse IFN‑ gamma Antibody.

Recombinant Mouse IFN-gamma (Catalog # 485-MI) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Inhibition of EMCV activity elicited by Recombinant Mouse IFN-gamma (2.5 ng/mL) is neutralized (green line) by increasing concentrations of Hamster Anti-Mouse IFN-gamma Monoclonal Antibody (Catalog # MAB4851). The ND₅₀ is typically 4-20 ng/mL.

IFN‑ gamma in Mouse Splenocytes.

IFN‑ gamma was detected in immersion fixed mouse splenocytes stimulated with calcium ionomycin and PMA (left panel) or unstimulated (right panel) using Hamster Anti-Mouse IFN‑ gamma Monoclonal Antibody (Catalog # MAB4851) at 15 µg/mL for 3 hours at room temperature. Cells were stained using an anti-hamster IgG secondary antibody (red) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of IFN-gamma by Flow Cytometry

Dependency on FOXP3 in IFN gamma -mediated PD-L1 expression. A, Expression of FOXP3 and PD-L1 in bladder cancer HT1376, T24, and SW780 cell lines without and with 100 ng/mL IFN gamma stimulation by flow cytometry. The mean with SD of at least three independent experiments, P values indicated. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. B, Expression of FOXP3 and PD-L1 in HT1376 CT and HT1376 FOXP3 KO bladder cancer cell lines without and with IFN gamma stimulation by qPCR. The mean of triplicates and representative of three independent experiments, P values indicated. C, Expression of FOXP3 and PD-L1 in HT1376 CT and HT1376 FOXP3 KO lines without and with IFN gamma stimulation by Western blotting. Sizes in kD as indicated. Sizes of FOXP3 delta 3 and FOXP3 depicted by overexpression in 293T cells. Full Western blots shown in Supplementary Fig. S1. D, Flow cytometry of FOXP3 and PD-L1 expression in HT1376 CT and HT1376 FOXP3 KO lines without and with IFN gamma stimulation. Representative of four independent experiments. E, Expression of FOXP3 and PD-L1 in rescue of a clonal HT1376 FOXP3 KO line by FOXP3 delta 3 and FOXP3 overexpression. Representative of three independent experiments. F, Expression of FOXP3 and PD-L1 in HT1376 FOXP3 KO lines with reconstitution of FOXP3 delta 3 and FOXP3 by Western blotting. Sizes in kD as indicated. Full Western blots shown in Supplementary Fig. S1. G, Flow cytometry of PD-L1 in HT1376 CT and HT1376 FOXP3 KO lines stimulated by IFN gamma without and with the presence of JAK1/2 inhibitor ruxolitinib (20 μmol/L), P values indicated. H, Expression of phosphorylated STAT1 (P-STAT1) in HT1376 CT and HT1376 FOXP3 KO lines stimulated by IFN gamma without and with the presence of JAK1/2 inhibitor ruxolitinib by Western blot. Full Western blots shown in Supplementary Fig. S1. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39099201), licensed under a CC-BY license. Not internally tested by R&D Systems.