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For testing, animals were anesthetized with ketamine (80 mg/kg) and xylazine (10 mg/kg) via i.p

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Immunostaining After organotypic culture or cryosection, the cochlear explants or tissue sections were fixed with 4% PFA, permeabilized with 1% TritonX-100 in PBS, and blocked by incubation in PBT-1 solution (0.1% Triton X-100, 8% donkey serum, 1% bovine serum albumin, and 0.02% sodium azide in PBS) at room temperature for 1 h

The downregulation of myeloid zinc finger 1 (MZF1) expression, which activates the Nrf2/GPX4 pathway and inhibits rankl-induced ferroptosis in osteoclasts during early cell differentiation, may be a key mechanism underlying ovarian removal (OVX) mice models of postmenopausal osteoporosis (37)

Primary antibodies against FOXO4 (Cell Signaling Technology 9472) and TP53BP1 (BD Biosciences 612523) were diluted in 1% BSA in PBST and incubated with the cells overnight at 4C