Quantitation ofIl1a,Tnf, andIl6mRNA bound to TTP-WT or TTP-NLSmutin RAW macrophages. TTP-WT, TTP-NLSmut, and TTP-TZFmut(negative control) expression was induced (0.5 g/mL dox, 24 h; DMSO: vehicle control) followed by treatment with 4sU (100 M, 7 h) and LPS (10 ng/mL, 6 h, added 1 h after 4sU). Bound RNA (pull-down) and total cellular RNA (input; 10% of the amount used for pull-down) were assessed by reverse transcription quantitative PCR (RT-qPCR), and fold differences were calculated (mRNA enrichment).36b4mRNA: non-TTP target control. IgG: mouse isotype control. Two-way analysis of variance (ANOVA), means of RT-qPCR data with SDs,n =3 (each in triplicates),*padj. < 0.01,****padj. < 0.0001.
