FIGURE 4.

Binding of WT-PEDF and EEE-PEDF to recombinant PNPLA2 and LR in in vitro binding assay. COS-7 cells were transfected with either GFP-PNPLA2 (A) or GFP-LR (B). Forty eight hours after transfection, cells were lysed, and recombinant proteins were immunoprecipitated (IP) using anti-GFP antibody. Thereafter, aliquots of washed immunoprecipitate-bound A/G beads were incubated with 4 μg of recombinant GST-WT-PEDF or GST-EEE-PEDF. Following incubation, the beads were boiled, and resolved protein complexes were analyzed by immunoblotting (IB) with anti-PEDF antibody. WCL, whole cell lysate. WT, WT-PEDF protein; EEE, EEE-PEDF protein.