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Cells were treated with ENDOs or vehicle and TLR9 activation in response to CpG (5 M) was analyzed by measuring SEAP activity, monitored using QUANTI-Blue (InvivoGene) following the manufacturers recommendations

The membranes were then incubated with the appropriate secondary antibodies (Santa Cruz Biotechnology, Inc.), conjugated with horseradish peroxidase (HRP) for 2 h at room temperature and rinsed three times with PBS

These results suggested that FSP1 overexpression lessened ferroptosis in CaOX-treated MTECs

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Cellular senescence in cancer: from mechanisms to detection