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. 2020 Feb 3;10(3):407–413. doi: 10.1002/2211-5463.12794

Figure 2.

Figure 2

PCBP2 binds to CREs in the 3′ UTR of sortilin mRNA. (A) RNA EMSA analysis. P, CRE probe mixed with binding buffer alone; N, whole‐cell macrophage lysates mixed with labeled WT probe and 50‐fold molar excess of nonspecific competitor; +IgG, negative control; +α‐P1, anti‐PCBP1 antibody supershift; +α‐P2, anti‐PCBP2 supershift; +α‐P1/P2, anti‐PCBP1 and anti‐PCBP2 supershift. (B) Sortilin mRNA coimmunoprecipitates with both PCBP1 and PCBP2. (C) RiboTrap analysis. Macrophage cytosol was mixed with the BrUTP‐labeled 3′ UTR synthesized in vitro. Protein/RNA complexes were precipitated with protein A/G magnetic beads bound to anti‐BrdU or rabbit IgG at 4 °C for 3 h. The BrUTP‐labeled RNA–protein complex was eluted with BrdU in PBS, followed by immunoblotting.