Figure 2.
Effect of AG-348 on the association of band 3 with ankyrin-1. (A-B) HbSS RBCs were treated with DMSO (Con) or 30-μM AG-348 in DMSO for different times (1, 2, 4, and 6 hours), and the level of membrane-associated ankyrin-1 was analyzed by western blotting and quantified by densitometry. The mean increase on ankyrin-1 of RBC treated with 30-μM AG-348 for 4 and 6 hours were 31% and 38%, respectively. ∗P ˂ .05. (C-D) HbSS RBCs were treated with DMSO (Con) or different concentrations (1, 3, 10, and 30 μM) of AG-348 in DMSO for 8 hours, and the interaction between ankyrin-1 and band 3 was measured by immunoprecipitation and quantified by densitometry analysis. The mean increase on the interaction between ankyrin-1 and band 3 of RBC treated with 1-, 3-, 10-, and 30-μM AG-348 were 5%, 38%, 45%, and 61%, respectively. ∗P ˂ .05. Three biological replicates of HbSS donors were used in all ex vivo assays.

Effect of AG-348 on the association of band 3 with ankyrin-1. (A-B) HbSS RBCs were treated with DMSO (Con) or 30-μM AG-348 in DMSO for different times (1, 2, 4, and 6 hours), and the level of membrane-associated ankyrin-1 was analyzed by western blotting and quantified by densitometry. The mean increase on ankyrin-1 of RBC treated with 30-μM AG-348 for 4 and 6 hours were 31% and 38%, respectively. ∗P ˂ .05. (C-D) HbSS RBCs were treated with DMSO (Con) or different concentrations (1, 3, 10, and 30 μM) of AG-348 in DMSO for 8 hours, and the interaction between ankyrin-1 and band 3 was measured by immunoprecipitation and quantified by densitometry analysis. The mean increase on the interaction between ankyrin-1 and band 3 of RBC treated with 1-, 3-, 10-, and 30-μM AG-348 were 5%, 38%, 45%, and 61%, respectively. ∗P ˂ .05. Three biological replicates of HbSS donors were used in all ex vivo assays.

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