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. Author manuscript; available in PMC: 2013 Nov 14.
Published in final edited form as: Int Immunopharmacol. 2011 Apr 13;11(9):10.1016/j.intimp.2011.03.021. doi: 10.1016/j.intimp.2011.03.021

Fig. 1.

Fig. 1

Reproducible CD154 induction among three different cell preparations in various stimulation conditions. Cells were plated in duplicate with various in vitro stimulations, and harvested for CD4 and CD154 staining after the indicated times of incubation. The non-stimulated group is representative of unfractionated splenocytes. (A) Either positively-selected CD4+ T cells or CD19-depleted splenocytes (“negatively-selected CD4+ T cells”) were cultured with plate-bound anti-CD3 and soluble anti-CD28 mAbs in vitro. When used, soluble IL-2 was added beginning at 72 h incubation. (B) CD19-depleted splenocytes were cultured alone or with plate-bound anti-CD3 and soluble anti-CD28 in vitro stimulation in three independent experiments. (C) Negatively-selected CD4+ T cells were stimulated in various conditions for CD154 induction: including with plate-bound anti-CD3, alone or in combination with soluble anti-CD28, removing the source of primary and/or secondary stimulation at 72 h, or supplementing with soluble IL-2 at 72 h. The data shown are representative of three independent experiments.