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Separated proteins were transferred onto Immobilon-P Transfer membranes (MilliporeSigma, IPVH00010), blocked for 1 h using 5% BSA in TBS0.5% Tween-20 and stained overnight with primary rabbit-anti-GCLC (Sigma-Aldrich, HPA036359) or affinity-purified rabbit-anti-GGT1 (GGT129, as previously described 63 ) 1:1,000 in BSA 2.5% overnight at 4 C

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The increased pulmonary flow through the intracardiac shunt results in increased shear stress on pulmonary endothelial cells, which then leads to vascular remodeling and dysfunction (Hawkins et al., 2011)

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