Skip to main content
. 2015 Dec 28;84(1):77–89. doi: 10.1128/IAI.01014-15

FIG 1.

FIG 1

Reactivity of the anti-LAG-3 MAb with LAG-3-expressing cells and bovine lymphocytes. (a) Flow cytometric analysis of bovine LAG-3. Cos-7 cells expressing LAG-3-EGFP (white area) and EGFP (shaded area) were stained with eight anti-LAG-3 MAb clones. Rat IgG1 (for 71-1A1, 71-1B3, 71-1H12, 71-2D8, 104-1G3, and 104-2F6), rat IgG2a (for 104-1F11), and rat IgG2b (for 104-2C9) were used as negative controls. (b) Western blotting of bovine LAG-3 protein from Cos-7 cells. Anti-LAG-3 MAb:71-2D8 recognized a LAG-3-EGFP protein band of ∼93 kDa. Anti-EGFP and antiactin antibodies were used as positive and loading controls, respectively. (c) Flow cytometric analysis of LAG-3 expression in CD4+ and CD8+ T cells. Freshly isolated bovine PBMCs were stained with anti-LAG-3:71-2D8 (white area), CD4, and CD8 MAbs. Rat IgG1 (gray area) was used as a negative-control stain. PBMCs were cultured with PBS (no stimulation) or PMA/ionomycin for 48 h and analyzed as described above.