MOESM5 of Cytokine production and phenotype of Histomonas meleagridis-specific T cells in the chicken

Additional file 5. IL-13 mRNA staining in HEK293T cells by PrimeFlowTM RNA Assay (Thermo Fisher Scientific). (A) Gating strategy for HEK293T cells in multicolor flow cytometry. After applying a time gate transfected cells were selected within a FSC-A/SSC-A plot followed by a dead cell exclusion gate using the Fixable Viability Dye eFluor® 506. Frequencies of IL-13 mRNA+ cells within live HEK293T cells were determined. (B) HEK293T cells were transfected with the pFLAG-CMV2 expression vector including a chicken IL-13 DNA insert (upper row) or a porcine IgE insert (lower row). Cells were stained with the IL-13 target probe and label probe (right panel) or with the label probe only (left panel). Percentages of IL-13 mRNA+ cells are indicated above the gate. Results are representative of two separate transfection experiments.