Figure 6.

Eftud2 regulates innate immunity by controlling the alternative mRNA splicing of MyD88. (A and B) Cells were treated with either Eftud2 siRNA or control (CT) nontargeting siRNA, were stimulated with 20 ng/ml LPS for 6 hr, and MyD88_L and MyD88_S mRNA levels were monitored by qPCR. (C–F) Cells were treated with the indicated siRNAs, were stimulated with 20 ng/ml LPS for 6 hr, and either IL-6 was monitored by ELISA (C) or Eftud2, MyD88_L, and MyD88_S mRNA levels were monitored by qPCR (D–F). mRNA levels were normalized such that 100% indicates mRNA in the presence of control RNAi in the absence of LPS. (G) Cells were exposed to either Eftud2 siRNA or control nontargeting siRNA (CT), were then treated with 20 ng/ml LPS for 6 hr, RNA was extracted, RT–PCR was performed to amplify the indicated mRNAs, and the amplified products were analyzed by agarose gel electrophoresis. Representative image is shown of PCR products using primers that bracket MyD88 exon 2 and that therefore simultaneously generate a 280-bp product corresponding to MyD88_L and a 145-bp product corresponding to MyD88_S.