FIGURE 3

OXTR Knockdown Decreases Proliferation and Activation of hiPSC-Derived Epicardial Cells. (A) Dose-response data for hEpiCs exposed to different concentrations of OXT over the course of 3 days, expressed as number of nuclei at each concentration; n = 10 per concentration. (B) Time course qRT-PCR data for OXTR in hEpiCs, dashed line corresponds to expression in undifferentiated hiPSCs. (C) Confocal immunofluorescent images for OXTR (green) and DAPI (blue) showing oxytocin receptor expression on the cell membrane and in the peri-nuclear region only in hEpiCs; scale bar: 25 µm. (D,E) Confocal immunofluorescent images (D) and quantification (E) of proliferating hEpiCs in both cell lines in the presence and absence of OXT at day 25 of differentiation. In (D), epicardial cells are labeled with WT1 (red), epithelial membranes are labeled with TJP1 (magenta), proliferating cells are labeled with Ki67 (green), nuclei are labeled with DAPI (blue); n = 10 images per condition, scale bar: 50 µm. (F) qRT-PCR data for scrambled and shOXTR hEpiCs, showing an increase in epicardial activation in control cells that is prevented after OXTR knockdown; n = 6 per condition, V: Vehicle.