Figure 3.

oxLDL-Induced DVL2 Activation is Dependent on WNT Secretion. (A) Representative western blots of THP-1-derived macrophages pre-treated with IWP-2 for 24 h at indicated concentrations. Preparation of THP-1 culture medium for western blot was described in the material and methods. In addition to protein estimation by bicinchoninic acid assay (BCA assay), Coomassie staining was used to confirm equal gel loading. IWP-2 demonstrated a concentration-dependent inhibition of WNT5A secretion into the cell culture supernatant, with no appreciable change in the intracellular content detected in whole-cell lysates. (B) Representative western blots of THP-1-derived macrophages pre-treated with vehicle (DMSO) or IWP-2 (5 μM) for 24 h and then treated with nLDL, or oxLDL (both 100 μg/mL) for an additional 24 h. IWP-2 inhibited oxLDL-induced DVL2 activation (black arrow), as evidenced by the reduction in intensity and electrophoretic mobility shift from ~90 to ~95 kDa. All experiments were performed in triplicate.