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Figure 2 | Cell Communication and Signaling

Figure 2

From: PAG/Cbp suppression reveals a contribution of CTLA-4 to setting the activation threshold in T cells

Figure 2

Decreased proliferation in primary human T cells. (A) Primary human T cells were transfected with siRNA oligonucleotide targeted against Renilla (ctrl) or PAG (PAGsi). Three days later, the cells were stimulated through CD3 plus CD28, lysed and immunoblotted with the indicated antibodies. Actin staining is shown as a loading control. The inset box shows the efficiency of PAG downregulation. (B) Primary human T cells were transfected with either Renilla siRNA (ctrl) or PAG siRNA (PAGsi) oligonucleotides and left for three days to downregulate PAG. Then, the cells were inoculated on a plastic plate coated with anti-CD3, anti-CD3+anti-CD28, anti-CD3 plus addition of exogenous IL-2 or uncoated (w/o). The incorporation of 3H-Thymidine was measured at 72 hours and is shown as the mean value of triplicates (±SD). Data were analyzed by the Student’s t-test (*, P<0.05; **, P<0.01; ***, P<0.001). (C) The cells transfected as in (B) were stimulated on an anti-CD3+anti-CD28 coated plastic plate for three days and the cell survival was assessed by annexin V-FITC and PI staining. Data are representative of three independent experiments. The phosphotyrosine blot in (A) was originally published in Blood: Smida, et al. 2007 [15]. Quantification of the blots can be found in the supplemental material (Additional file 3: Figure S3).

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