Anti ifn γ apc clone xmg1

Mouse splenocytes were stimulated ex vivo with SFTSV OLP pools. Brefeldin A (GolgiPlug, BD Biosciences) and monensin (GolgiStop, BD Biosciences) were added to the culture 1 h after stimulation and the culture was maintained for 5 h. The Live/Dead Fixable Red Dead Cell stain kit (Invitrogen), anti-CD19–PE-CF594 (clone 1D3, BD Biosciences, 562291, 1:100), anti-CD3–BV510 (clone 145-2C11, BD Biosciences, 563024, 1:100), anti-CD4–Alexa Flour 700 (clone RM4-5, BD Biosciences, 557956, 1:100), and anti-CD8–APC-H7 (clone 53-6.7, BD Biosciences, 560182, 1:100) were used for immunostaining at 4 °C for 20 min. For intracellular staining, cells were permeabilized and fixed using the BD Cytofix/Cytoperm kit (BD Biosciences) at 4 °C for 20 min, which was followed by staining with anti-IFN-γ–APC (clone XMG1.2, BioLegend, 505809, 1:100), anti-TNF–PE (clone MP6-XT22, BD Biosciences, 554419, 1:100), and anti-IL-2–PE-Cy7 (clone JES6-5H4, BD Biosciences, 560538, 1:100) at 4 °C for 20 min. All data were collected using a LSRΙΙ flow cytometer (BD Biosciences), and Boolean gating was performed using FlowJo software to analyze the polyfunctionality of T cells. Charts were visualized using SPICE software (https://niaid.github.io/spice/).