Figure 5.

Flow cytometry results of annexin-V and propidium iodide (PI) staining of apoptotic cells following the GANT61 treatment of B16 cells. Each population was hierarchically linked as P4 to P5 to P6 of gating. Early apoptosis was defined as PI-negative and annexin V-positive, and late apoptosis was defined as PI-positive and annexin V-positive. Furthermore, PD-L1-positive cells were counted among those cells via gating. (A) Early apoptosis: no significant difference in the number of cells stained as PI- negative and annexin V-positive is observed between the control and GANT61 groups (χ²-test, p = 0.058, n = 4). No significant difference in the number of cells stained with PD-L1-positive among early apoptotic cells is detected between the control and GANT61 groups (χ²-test, p = 0.134, n = 4). (B) Late apoptosis: a significant increase in the number of cells stained as PI-positive and annexin V-positive is observed in the GANT61 group compared to the control group (χ²-test, ** p = 0.000, n = 4). A significant decrease in the number of cells stained with PD-L1-positive among late apoptotic cells is detected in the GANT61-treated group compared to the control group (χ²-test, ** p = 0.000, n = 4).