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Whole-mount confocal staining Differentiated urothelia on transwells were washed and fixed in PBS with 4% paraformaldehyde for 15 min and rinsed 3 times with PBS

Westerterp-Plantenga MS, Lemmens SG, Westerterp KR

Fe 3+ is reduced to Fe 2+ in the intestinal lumen by iron-reducing enzymes, including duodenal cytochrome b reductase (Dcytb), and transported to the enterocytes via the divalent metal 1 transporter (DMT1)

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Thiols quantification Because thiols can impact protein function by the formation of intra- and inter-molecular disulfide bond(s) with proteins 59 , or may exist as free thiols, we measured both total and free thiol levels

This process effectively improved mitochondrial dysfunction by maintaining mitochondrial membrane potential, reducing mitochondrial swelling, and inhibiting excessive ROS production, thereby alleviating RGCs apoptosis and axonal degeneration [58]