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Because the reaction of NO with free thiols competes with the same substrate (e.g., GSH), the excessive NO formation may further lead to significant depletion of GSH in SZ
For iHLO differentiation, iPSCs were dissociated with Versene (Thermo Fisher Scientific, catalog: 15040066) on Day 0 and plated into an AggreWell plate (STEMCELL Technologies, catalog: #34421) in Stemflex Medium (Thermo Fisher Scientific) to form aggregates, which were then transferred to an ultralow attachment dish on Day 1 and cultured in RPMI1640 supplemented with B27 supplement minus insulin (RPMI/B27-Insulin), CHIR99021 (1 M), Activin A (100 ng/mL), WNT3a (50 ng/mL), and Rock Inhibitor (Y27632, 5 M)
Young IS, Woodside JV