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For in vitro exflagellation analysis, 2.5 l of tail vein blood was collected in the exflagellation medium of RPMI-1640 pH 8.0 containing 100 M xanthurenic acid (Sigma-Aldrich, D120804) and 10% heat-inactivated FBS and incubated at 19 C for ~12 min, followed by immediate counting of exflagellation centers by light microscopy

In autoimmune diseases, mitochondrial dysfunction and impaired mitophagy fuel chronic inflammation by releasing ROS and mtDNA, which activate innate immune pathways and promote autoreactive immune responses

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