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Day 10 cultured SCD erythroblasts were treated with 75 M hemin (#51280, Sigma) (dissolved in 0.25 M NaOH then neutralized with HCL to pH7.4), 50 M dimethyl fumarate (#242926, Sigma) (dissolved in DMSO), 0.5 mM (2S)-Tetrahydro-5-oxo-2-furancarboxylic acid (TFMB-L2HG) (#SML2563, Sigma) (dissolved in DMSO), 0.5 mM S-(5-Adenosyl)-L-methionine iodide (SAM) (#A4377, Sigma) (dissolved in water), and cultured under hypoxia (1% O 2 ) using Heracell 150i incubator equipped with an Oxygen (O 2 ) sensor and Nitrogen (N2) gas supply and analyzed on day 12 of culture
Pelleted cells were sectioned in a RMC Boeckeler ultramicrotome using a Diatome diamond knife at a thickness setting of 60 nm
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Massafra et al (21, 22) reported that a significant positive correlation existed between physiological estrogen and antioxidant expression during the menstrual cycle, but no significant changes occurred in catalase