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Fig. 4 | Journal of Inflammation

Fig. 4

From: Functional regulation of Zfp36l1 and Zfp36l2 in response to lipopolysaccharide in mouse RAW264.7 macrophages

Fig. 4

Zfp36l1 and Zfp36l2 interact with Mkp-1 3′UTR in RAW264.7 cells. a Interaction of TTP family proteins and Mkp-1 mRNA. Biotinylated-Mkp-1 3′UTR fragments were incubated with cytosolic extracts from control RAW264.7 or LPS-treated cells for 15, 30, 60, or 120 min. The biotinylated RNAs and associated proteins were precipitated by streptavidin beads, separated by SDS-PAGE, and analyzed by western blotting with anti-Ttp, anti-Zfp36l1, anti-Zfp36l2, and anti-Caf1a as indicated. Biotinylated 18S rRNA was used as a negative control. The pull-down protein levels were quantified as indicated below the lanes. b Interaction of Zfp36l1 with 14-3-3. E. coli–expressed GST-14-3-3 ζ or GST was bound on glutathione Sepharose 4B. The beads were incubated with the cell lysates from LPS-stimulated RAW264.7 cells for 0, 15, 30, 60, or 120 min. The pulled-down protein complexes were separated by SDS-PAGE and analyzed by western blotting with anti-Zfp36l1. All of the experiments were performed at least three times, and representative data were displayed

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