Intracellular expression of IL-17A and IFN-γ by activated bovine T cell subsets. PBMC from four cattle were stimulated with phorbol 12-myristate 13 acetate, ionomycin and brefeldin A in RPMI culture medium for 4 h. Cells were harvested and stained for viability and with mabs specific for cell-surface phenotypic markers and intracellular cytokines as described in Table 3 and “Expression of intracellular IL-17A and IFN-γ by bovine and ovine T cell subsets section”. Cells were stained for CD4 with mab CC8-PE at 1:20 dilution (A, D), for CD8β with mab CC58-PE at 1:20 dilution (B, D) and for WC-1 (γδ T cells) with mab CC15-PE at 1:200 (C, E). Intracellular cytokine staining for IL-17A was conducted using mab eBioDEC17-APC at a 1:20 dilution (A–C) and for IFN-γ using mab CC302-Alexafluor 647 at a 1:200 dilution (D–F). Data are shown for PBMC from one representative animal of four.