Fig. 3

Zfp36l1 and Zfp36l2 destabilize Mkp-1 mRNA in resting RAW264.7 cells. a shLuc, shL1, shL2, shL1 + L2 represent luciferase knockdown cells, Zfp36l1-knockdown cells, Zfp36l2-knockdown cells, and dual Zfp36l1- and Zfp36l2-knockdown cells, respectively. The upper two panels show the knockdown efficiency. β-tubulin was used as a loading control. Whole-cell extracts were collected for western blotting analysis using the indicated antibodies. b Basal levels of Mkp-1 mRNA were detected by quantitative PCR in different knockdown cells. c Analysis of Mkp-1 mRNA half-life in different knockdown cells. Actinomycin D (10 μg · mL^–1) was added to stop transcription for 0, 0.5, 1 or 2 h. The remaining mRNA was detected by quantitative PCR. Mkp-1 mRNA half-life was calculated by exponential regression, 19 min in control cells and 91, 96 and 95 min in Zfp36l1, Zfp36l2 and dual knockdown cells, respectively. d Basal levels of Ttp, Tnfα, Icam-1, and Ccl-2 mRNAs were examined by quantitative PCR in different knockdown cells and normalized to the shLuc control. All of experiments were independently performed at least three times