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. 2011 Oct 27;6(10):e27020. doi: 10.1371/journal.pone.0027020

Figure 3. Calpastatin-overexpression in Th cells resulted in decreased IL-6 and IL-17 production.

Figure 3

(A) Naïve CD4+ T cells were cultured under neutral, Th1, Th2, or Th17 conditions. On day 1, these cells were infected with mock-GRV or calpain- or CS-dIV-GRV and expanded on day 2 with IL-2. On day 4 (Th17 conditions) or on day 6 (neutral, Th1, and Th2 conditions), the cells were restimulated with PMA/ionomycin for ICS. Representative results for GFP+-gated cells are shown (n = 11 for IFNγ, n = 10 for IL-4, n = 6 for IL-6 and IL-17). (B) Statistical analysis of the results in panel A by Steel's test. (C) Naïve CD4+ T cells were cultured under neutral conditions and infected with mock-GRV or CS-dIV-GRV. On day 6, these cells were restimulated with PMA/ionomycin for FACS. Representative data of GFP+, GFP−-gated cells are shown in the left panel, and the statistical analysis is shown in the right panel (n = 3) (D) Retrovirus-infected Th cells were stained with PKH-26 dye and then stimulated with anti-CD3/28 monoclonal antibodies. The fluorescence intensity of the PKH-26 dye in infected (GFP+) and non-infected (GFP−) cells was analyzed by FACS. *p<0.05 vs. mock.