Figure 3.

The effect of CD39/CD73 inhibiting on the apoptosis and mitochondrial membrane potential (ΔΨm) of dNK cells. dNK cells were isolated from healthy decidua of first-trimester pregnancies (n = 3), incubated in the absence or presence of the CD39 inhibitor ARL67156 for 12 hours (A) or 18 hours (B), and then double stained with Annexin V and DAPI (mean ± SEM, ***P < 0.001, two-tailed t tests). (C) Microscope images taken from the JC-1 monomer fluorescence channel (green) and aggregate fluorescence channel (red) of dNK cells (n=5) with and without incubation with ARL67156 for 12 h. The monomeric JC-1 form was excited using a 525 nm laser, observed at an emission wavelengths of 514~529 nm, and is shown in green. The aggregate form was excited using a 566 nm laser, observed at 585~590 nm, and is shown in red. (D) Flow cytometry-based JC-1 assay as a measure of changes in mitochondrial membrane potential in dNK cells (n=5) induced by the CD39 inhibitor. The upper left quadrant indicates the cells with more JC-1 as aggregates with red fluorescence (i.e., normal ΔΨm), the lower right quadrant indicates the cells with more JC-1 as monomers with green fluorescence (i.e. low ΔΨm). (mean ± SEM, *P < 0.05 compared with the untreated group).