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. 2012 Dec 17;209(13):2383–2394. doi: 10.1084/jem.20112631

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© 2012 Nishida et al.

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Figure 8. — Requirement of PKCθ for CAG-mediated proliferation. (A-C) Purified colonic CD4⁺ T cells from βIL-2 DKO mice with reconstitution of WT-derived (WT>βIL2) versus T/C2GnT tg–derived (C2GnT>βIL2) memory CD4⁺ T cells were stimulated with anti-CD3/CD28–coated beads, and after removal of beads they were cultured again in the presence or absence of galectin-4 for 1 (A) and 3 h (B). They were pulsed with BrdU 1 h before the analysis and subjected to flow cytometric analysis. The percentage of BrdU⁺ cells is summarized in C. *, P < 0.01 (n = 4–7). (D) Purified colonic CD4⁺ T cells from βIL-2 DKO mice with reconstitution of WT-derived (WT>βIL2) versus T/C2GnT tg–derived (C2GnT>βIL2) memory CD4⁺ T cells were pretreated with KT5720 (left) or rotterin (right), and they were then stimulated with anti-CD3/CD28–coated beads. After removal of beads, they were cultured in the presence of galectin-4 for 3 h, with 1-h pulse with BrdU, and subjected to flow cytometric analysis for evaluation of BrdU incorporation (top) and galectin-4 binding (bottom). Data shown are one representation of two independent experiments. (E and F) Purified CD4⁺CD45RB^low memory T cells from the spleen of GFP Tg versus PKCθ^−/− mice were co-transferred into βIL-2 DKO mice. The intensity of galectin-4 binding on (E) and the proliferative response of (F) reconstituted GFP⁺ CD4⁺ (PKCθ-intact) versus GFP⁻ CD4⁺ (PKCθ-deficient) cells in the same recipient colon are shown. The mean (n = 4) of percentages of BrdU⁺ cells in GFP⁺ (PKCθ-intact) versus GFP⁻ (PKCθ-deficient) CD4⁺ T cells are summarized in G. *, P < 0.01.