Anti cd8a ct 8

Isolated LP cells were analyzed using FACS Canto II (BD, Franklin Lakes, NJ, USA). Dead cells were excluded using Live/Dead fixable dead cell stain (Thermo Fisher Scientific, Waltham, MA, USA). The following anti-chicken antibodies were used for staining: anti-CD3 (CT-3; Southern Biotech, Birmingham, AL, USA), anti-CD4 (CT-4; Southern Biotech, Birmingham, AL, USA), anti-CD8a (CT-8; Southern Biotech, Birmingham, AL, USA), anti-TCR γδ (TCR-1; Southern Biotech, Birmingham, AL, USA), anti-MHC II (2G11; Southern-Biotech, Birmingham, AL, USA), anti-Bu-1 (AV20; Southern Biotech, Birmingham, AL, USA), and anti-Monocyte/Macrophage (KUL01; Southern Biotech, Birmingham, AL, USA). All antibodies were diluted 1:200 in PBS and incubated for 30 min under dark conditions. Then, all samples were fixed using 4% paraformaldehyde (PFA) and stored at 4 °C until analysis. The analysis was conducted by two panels: (1) MHC II, Bu-1, and monocyte/macrophage for B cells and APCs; (2) CD3, CD4, CD8a, and TCR γδ for T cells. For the detailed gating strategy of flow cytometry analysis, refer to Supplementary Figure S1.

Isolated LP cells were analyzed using FACS Canto II (BD, Franklin Lakes, NJ, USA). Dead cells were excluded using Live/Dead fixable dead cell stain (Thermo Fisher Scientific, Waltham, MA, USA). The following anti-chicken antibodies were used for staining: anti-CD3 (CT-3; SouthernBiotech, Birmingham, AL, USA), anti-CD4 (CT-4; SouthernBiotech, Birmingham, AL, USA), anti-CD8a (CT-8; SouthernBiotech, Birmingham, AL, USA), anti-TCRγδ (TCR-1; SouthernBiotech, Birmingham, AL, USA), anti-MHC II (2G11; SouthernBiotech, Birmingham, AL, USA), anti-Bu-1 (AV20; SouthernBiotech, Birmingham, AL, USA), and anti-Monocyte/Macrophage (KUL01; SouthernBiotech, Birmingham, AL, USA). All antibodies were diluted 1:200 in PBS and incubated for 30 min under dark conditions. Then, all samples were fixed using 4% paraformaldehyde (PFA) and stored at 4 °C until analysis. The analysis was conducted by two panels: (1) MHC II, Bu-1, and monocyte/macrophage for B cells and APCs; (2) CD3, CD4, CD8a, and TCR γδ for T cells.