Fig. 5.

AG1 reduces hemolysis upon exposure to oxidative stressors. a AG1 reduced the extent of hemolysis of 5% erythrocyte suspension treated with either 1 mM chloroquine (CQ) for 4 h under light or 1 mM diamide for 4 h at 37 °C (n = 7 independent blood samples, ^*p = 0.0372, ^**p = 0.0019, ^****p < 0.0001, one-way ANOVA). b–d AG1 significantly increased GSH levels and reduced ROS levels, when 5% erythrocyte suspension was exposed to either 1 mM chloroquine or diamide for 3 h at 37 °C, which was consistent with increased G6PD activity (n = 9–11, ^*p < 0.05, ^**p < 0.01, ^****p < 0.0001, one-way ANOVA). e Band 3 protein was clustered (cBand3) when 5% erythrocyte suspension was treated with chloroquine, which was alleviated by AG1 treatment. Each lane represents one individual sample, and quantification is provided in Supplementary Fig. 7a. f–h AG1 (1 μM) improved storage of erythrocytes (5% suspension) at refrigerated temperature by reducing hemolysis and concomitant protein leakage for 28 days (n = 13 independent blood samples, ^*p < 0.05, two-tailed unpaired Student’s t-test), which corresponded with increased G6PD activity (n = 4, ^*p = 0.0323, ^***p = 0.0003, one-way ANOVA). Each sample was re-treated with AG1 every week. Representative hemolysis phenotypic images are provided below f. Error bars represent mean ± SD. NT: no treatment, CQ: chloroquine, cBand3: clustered band 3 protein