Fig. 4.

Spt6 knockdown inhibited CSR in the artificial switch substrate SCI (μ, α) in NIH 3T3 cells. (A) NIH 3T3 cells expressing mAID-ER and SCI (μ, α) were introduced with d-siRNAs against mSpt6, mTrim28, mAID, and LacZ together with a DsRed-expressing plasmid as a transfection indicator. Twenty-four hours after transfection, cells were stimulated with OHT for 36 h. The percentages of switched CD8α⁺ cells in the live and DsRed⁺ population are indicated. Mean ± SD values were obtained from triplicate experiments. (B) Quantitative PCR analyses for pre- (Pre-Tr1 and Pre-Tr2) and postswitch transcripts (Post-Tr), AID-ER, Spt6, and Trim28 transcripts. CYPH, cyclophilin.