Autophagy Deficiency Leads to Impaired Antioxidant Defense via p62-FOXO1/3 Axis

Figure 1

Mitochondrial dysfunction, increased ROS, and impairment of antioxidant enzymes occur in response to chemical inhibitors of autophagy. (a, b, d, and e) HEK293T cells were treated with autophagy inhibitors Bafi A1 (100 nM), CQ (50 μM), or 3-MA (10 mM) for 24 hours or with 10 mM 3-MA in a time course within 72 hours before harvest. Then, mitochondrial DNA (as D-loop DNA) copy number was detected by real-time PCR () (a), mitochondrial membrane potential (MMP) was detected with the JC-1 fluorescent probe () (b), ROS level alterations were determined by a fluorescence microplate reader after staining with DCFDA () (d), and protein levels of antioxidant enzymes were determined by W.B. (e). (c and f) WT MEF cells were treated with CQ at indicated concentrations for 24 hours or with 50 μM CQ for the indicated time period before harvest, then oxygen consumption rate (OCR) was determined using the Seahorse XF24 Analyzer () (c) and mRNA levels of antioxidant enzymes were determined by q-RT-PCR () (f). Values are represented as . ; ; ; vs. control group.

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