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Enhanced green fluorescent protein (EGFP)-tagged wild type paxillin and Y31F, Y118F mutants were prepared as described previously13. ROS biosensors targeted to cytosol, mitochondria, and nucleus were obtained from Evrogen (Moscow, Russia). Constructs were added to HLMVECs grown to ∼ 80% confluence in EBM-2-MV growth media (Lonza) supplemented with 10% FBS. After overnight culture, the medium was replaced with fresh complete medium and cells were cultured for additional 48 h before experimentation.
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