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Figure 1 | Veterinary Research

Figure 1

From: Decreased STEC shedding by cattle following passive and active vaccination based on recombinant Escherichia coli Shiga toxoids

Figure 1

Gating strategy used to quantify CD25 expression on inating vitro re-stimulated CD4+CD45RO+ and CD8αhiCD45RO+ cells by flow cytometry. Representative dot plots obtained with PBMC after 5 days of in vitro cultivation with Concanavalin A and subsequent tri-colour immunolabelling. A In a first analytical step, the non-blast cell (Region R0) and blast cell population (R1) were acquired for gating based on its forward versus side scatter (FSC/SSC) characteristics. B In samples labelled with anti-bovine CD4 antibodies, CD4+ cells within the blast cell population were defined by R2 as such that less than 2% of events were considered positive in the corresponding isotype control sample. C In samples labelled with anti-bovine CD8α antibodies, CD8αhi cells were defined by R3, thereby excluding the majority of CD8αlo cells. D Cells within R2 (CD4+) and R3 (CD8αhi) were displayed on CD45RO/CD25 dot plots and the geometric mean for detection of CD25 expression was assessed for all CD45RO+ within the CD4+ and CD8αhi, respectively, as defined by R4 (shaded area).

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