Figure 3
From: PAG/Cbp suppression reveals a contribution of CTLA-4 to setting the activation threshold in T cells
PAG down-regulation results in unresponsiveness. (A) Primary human T cells transfected either with Renilla (ctrl) or PAG (PAGsi) siRNA were stimulated on an anti-CD3 coated plastic plate for 24 hours and the production of IL-2 was assessed by intracellular FACS staining. (B) The cells treated as in (A) were used to determine the upregulation of surface CD25 expression by flow cytometry. In the histograms, white bars represent ctrl, grey bars PAGsi. (*, P<0.05; **, P<0.01) (C) Primary human T cells transfected either with Renilla (ctrl) or PAG (PAGsi) siRNA were stimulated with CD3+CD28 antibodies for the indicated time, lysed and subjected to Western blotting. The activation of ZAP70 and PLCγ was determined by phospho-specific antibodies. Actin staining is shown for equal loading. Data are representative of three (A, B) or two (C) independent experiments. Quantification is shown in the supplemental material (Additional file 6: Figure S5).