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Cells were then fixed with 4% paraformaldehyde, treated with 0.1% Triton X-100 in PBS for 10 min, incubated with 0.1% Tween 20, 0.3 M glycine, and 1% BSA in PBS for 2 h and incubated overnight at +4 C with monoclonal antibodies against -III tubulin conjugated to Alexa Fluor 647 (1:500, 657405, Biolegend, USA) in the same buffer

The impact of hypoxia on cytosolic NADP redox dynamics had remained less clear, since lack of oxygen as an electron sink causes metabolic reduction, but also boosts H 2 O 2 production and glutathione oxidation in different phases of the hypoxic episode [37]

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Afterward, the sections were incubated for 2 h at room temperature in the dark with secondary antibodies: Cy3-labeled goat anti-rabbit (1:200, Jackson ImmunoResearch) and 488-labeled goat anti-mouse (1:200, Jackson ImmunoResearch)

It also helps prevent inflammation and disease