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. 2005 Dec;167(6):1661–1675. doi: 10.1016/S0002-9440(10)61249-7

Figure 1.

Figure 1

Characterization of Th1 and Th2 cells from wild-type and PSGL-1-deficient DO11.10 TCR transgenic mice. Naïve CD4+ T cells from wild-type or PSGL-1-deficient DO11.10 mice were cultured with irradiated BALB/c splenocytes and OVAp for 7 days (Th1) or 14 days (Th2) under Th1 (rmIL-12 and anti-IL-4) or Th2 (rmIL-4, anti-IL-12, and anti-IFN-γ) polarizing conditions. A: For detection of intracellular cytokine expression 1 × 106 cells per condition were restimulated with PMA and ionomycin for 6 hours. Monensin was added for the final 4 hours to block cytokine secretion. T cells were fixed, permeabilized, and stained intracellularly for IFN-γ and IL-4. The frequency of IFN-γ+ and IL-4+ cells was assessed by flow cytometry on the lymphoid gate. Quadrant percentiles of lymphocyte-gated cells are indicated. B: For detection of surface phenotype, polarized Th1 and Th2 cells derived from wild-type (solid line) and PSGL-1-deficient (dotted line) DO11.10 mice were stained with monoclonal antibodies for the indicated adhesion molecules and analyzed by flow cytometry. Plots represent populations in the lymphocyte gate (30,000 events).