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. 2020 Mar 15;10(10):4453–4465. doi: 10.7150/thno.42354

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CREB activation is essential in PACAP-mediated hepatic autophagy in liver IRI. PACAP+DMSO, PACAP+CREB inhibitor or PBS pre-treated WT mice were subjected to partial liver warm ischemia (90 min). Ischemic liver tissues were harvested at 6 h of reperfusion for (A) western blot of Atg5, LC3 I and LC3 II; (B) qPCR-assisted detection of LC3a, LC3b, Beclin-1, EGF, HGF, and c-Met (n=4-6/group, *p<0.001). (C) Immunofluorescence staining: LC3 (green), F-actin (red) and DAPI (blue): LC3 accumulation in cytoplasm and nuclear (magnification: x400, scale bar represents 20µm); (D) PI staining (red): live cells (blue arrows) and dead cells (red arrows) (magnification: x200, scale bar represents 50µm); (E) supernatant ALT and LDH levels (*p<0.001, n=6/group).