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. 2012 Aug 1;7(8):e41972. doi: 10.1371/journal.pone.0041972

Figure 1. CD47 status is differentially regulated on TCR- activated human CD4 T cell subsets.

Figure 1

(A–B) CFSE-labeled TN and TCM cells were stimulated with immobilized anti-CD3 and soluble anti-CD28 mAbs for 6 days. (A) CD47 (using human SIRP-α-Fc protein or anti-CD47 mAb, clone B6H12) and CCR7 expression was analyzed by flow cytometry. (B) Phenotype of divided CD47low and undivided CD47high cells at day 6 of TN cultures. (C) Strategies to examine CD47 expression on ex vivo isolated human T cells gated on CD4+ T cells. (D) CD47 expression on CD4 T cell subsets using SIRP-α-Fc and anti-CD47 antibodies (B6H12 and 2D3). The mean ± standard deviation (SD) for 16 donors is shown (Anova test: ***p<0.0001). (E) Western blot analysis for CD47 protein on whole-cell lysates using 2D3 mAb. (F) Confocal immunofluorescence of CD47 using SIRP-α-Fc or anti-CD47 (B6H12) antibodies. (A–C; E and F) Data are representative of 3 to 6 independent experiments.