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The protein was transferred to 0.2 m pore-sized nitrocellulose (wet transfer, 1 hour, 100 mA, RT) and then blocked with 5% non-fat dry milk in TBS-T (20 mmol/L TrisHCl, 150 mmol/L NaCl, 0.05% Tween 20, adjusted to pH 7.4) and incubated with the following primary monoclonal antibodies: anti--actin, anti-cleaved-PARP, anti-cleaved Caspase-3, anti-cleaved Caspase-9, and anti-COX IV (for apoptosis evaluation) or anti-LC3A/B, anti-Beclin-1, anti-Atg12, and anti-Atg-7 (for autophagy evaluation) antibodies, diluted 1:1,000 in blocking buffer overnight at 4C

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