Figure 5.

WNT5A Receptors, FZD5 and ROR2, are Co-localized in vitro in Response to oxLDL and WNT5A. Representative confocal immunofluorescence (IF) images of THP-1-derived macrophages. Cells were differentiated with PMA for 24 h and then treated with rWNT5A (100 ng/mL), (nLDL) or oxLDL (both 100 μg/mL) for an additional 24 h. Cells were then fixed, dual-stained with FZD5 (Red) and ROR2 (Green), and counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Compared to untreated cells (UT), rWNT5A increased the co-localization of the FZD5 and ROR2 receptors at the plasma membrane (Pearson's Correlation: UT = 0.548, rWNT5A = 0.78). Further, oxLDL-treated cells showed an even greater increase in co-localization of the FZD5 and ROR2 receptors compared to all groups (Pearson's Correlation: oxLDL = 0.875). Immunofluorescent images are at 60x with equally zoomed insets for clarity (scale bar= 20 μm). Experiments were performed in duplicate and repeated three independent times. Representative negative control and isotype control images are included in the data supplement (Supplementary Figure 2).