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. Author manuscript; available in PMC: 2014 Jul 7.
Published in final edited form as: J Immunol. 2014 Jun 13;193(2):889–900. doi: 10.4049/jimmunol.1303389

Figure 3.

Figure 3

(A) In vitro activation of CD4+ T cells induces secretion of sCTLA-4. CD4+ T cells of four healthy donors were stimulated in vitro with anti-CD3/CD2 mAb and culture supernatants harvested at the indicated time points. CD4+ T cells (3 × 106 cells/ml) from the four donors were cultured on separate days. Isotype controls were negative for all samples (not shown). (B) Secretion of sCTLA-4 by CD4+ T cells (1 × 106 cells/ml) of three independent donors stimulated for 72 hours in vitro with either anti-CD3/CD2 mAb or anti-CD3/CD28 microbeads. (C) Detection of sCTLA-4 following immunoprecipitation and Western blot analysis. sCTLA-4 protein in culture supernatants from activated CD4+ T cells and from HeLa cells transfected with sCTLA-4 were immunoprecipitated using 4B8 mAb followed by 4017 polyclonal Ab immunoblotting. No sCTLA-4 was immunoprecipitated with the corresponding IgG2b isotype control (not shown). Western blot gel is representative of more than three independent experiments.