Knowkdown using siRNA in CD4+ T cells were performed as described by Ciofani et al.27 (link). CD4+ T cells from wild-type mice were cultured for 16 h under Th0 conditions, and 300 pmol of control or c-Jun siRNA (Invitrogen) was introduced using the Mouse T cell Nucleofector Kit (Lonza) according the manufacture’s instruction. After recovery by incubation in fully supplemented culture media (Lonza), the cells were cultured for 3 days under Th17 conditions. Sequences for c-Jun siRNAs are as follows: #1, 5′-UACUGUAGCCGUAGGCACCGCUCUC-3′; #2, 5′-AUGACUUUCUGCUUAAGCUGUGCCA-3′; #3, 5′-AAACGUUUGCAACUGCUGCGUUAGC-3′.
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