Figure 4.

VIT-2763 alone or in combination with DFX improved the functional parameters of RBCs in Hbb^th3/+ mice. RBCs were gated as mature RBCs (Ter119^hiCD71^neg) and analyzed for reactive oxygen species (ROS) (CM-H₂DCFDA), apoptosis (annexin V as staining for phosphatidylserine (PS)), and retention of mitochondria (MitoTracker Deep Red). VIT-2763 alone or in combination with DFX reduced the percentage of ROS-positive RBCs (a), lowered PS exposure on peripheral RBCs (b), and improved the elimination of mitochondria in RBCs (c) of Hbb^th3/+ mice compared to Hbb^th3/+ mice treated with the vehicle or DFX alone. Results represent the mean ± SD. Statistical significance was determined using one-way ANOVA with Dunnett’s multiple-comparison test. * p < 0.05, ** p < 0.01, and *** p < 0.001. Wild-type (WT) littermates received both vehicles and were included as controls. n = 9–13 mice per group.