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

Figure 4. Silencing of the calpain small subunit suppressed the IL-17 production from Th cells.

Figure 4

(A) Naïve CD4+ T cells were cultured under neutral conditions, and treated with siRNA against calpain small subunit (CPNS). This experiment was carried out using two siRNAs (CPNS1; C1 and CPNS2; C2). On day 3 and 7, these cells were lysed and subjected to western blot analysis. NC; negative control. (B) On day 6, the cells in panel A were washed 3 times and restimulated with an anti-CD3 antibody for 48 hours. The culture supernatants were then collected, and IL-17 was measured by ELISA. Statistical analysis of eight independent experiments is shown (Steel's test). (C) On day 6, the cells in panel A were incubated with X-VIVO™ 20 (without FCS) for the last 6 hours, then lysed and subjected to western blot analysis with the indicated antibodies. Similar results were obtained in three independent experiments. The band densities of pSTAT5 were measured and normalized to that of total STAT5, then the percent change from NC was calculated. Statistical analysis of three independent experiments is shown in the right panel. *p<0.05 vs. NC.