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miR-30b inhibits autophagy to alleviate hepatic ischemia-reperfusion injury via decreasing the Atg12-Atg5 conjugate

Li, Shi-Peng·2016·World Journal of Gastroenterology
DOI 10.3748/wjg.v22.i18.4501
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Figure 3 miR-30b inhibits autophagy by sequestering Atg12 in AML12 cells. A and B: The predicted miR-30b binding site on the Atg12 mRNA 3′-untranslated region (UTR) is shown, and a luciferase reporter assay was performed to determine the effects of miR-30b on the Atg12 mRNA 3′-UTR; C: Western blot was used to detect expression of Atg12 and LC3 in AML12 cells treated with miR-30b mimics or inhibitor; D: Confocal immunofluorescence of AML12 cells demonstrated increased numbers of GFP-RFP-LC3 dots in the IR group; when autophagy is induced, both GFP and RFP are expressed as yellow dots (autophagosomes). Red dots represent autolysosomes as the GFP degrades in an acid environment. Scale bars = 20 μ m; E: TEM images of AML12 cells after ischemia followed by reperfusion at 12 h. TEM images show representative examples of autophagosomes (red arrows). Scale bars = 1.0 μ m. And the data were quantified by counting the number of autophagosomes per cross-sectioned cell; a P < 0.05, b P < 0.01, c P < 0.001 vs miR-NC group. Every experiment was repeated three times.
Figure 3 miR-30b inhibits autophagy by sequestering Atg12 in AML12 cells. A and B: The predicted miR-30b binding site on the Atg12 mRNA 3′-untranslated region (UTR) is shown, and a luciferase reporter assay was performed to determine the effects of miR-30b on the Atg12 mRNA 3′-UTR; C: Western blot was used to detect expression of Atg12 and LC3 in AML12 cells treated with miR-30b mimics or inhibitor; D: Confocal immunofluorescence of AML12 cells demonstrated increased numbers of GFP-RFP-LC3 dots in the IR group; when autophagy is induced, both GFP and RFP are expressed as yellow dots (autophagosomes). Red dots represent autolysosomes as the GFP degrades in an acid environment. Scale bars = 20 μ m; E: TEM images of AML12 cells after ischemia followed by reperfusion at 12 h. TEM images show representative examples of autophagosomes (red arrows). Scale bars = 1.0 μ m. And the data were quantified by counting the number of autophagosomes per cross-sectioned cell; a P < 0.05, b P < 0.01, c P < 0.001 vs miR-NC group. Every experiment was repeated three times.CC-BY-NC · From the PDF
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Figure 5 Model of miR-30b inhibition of autophagy to alleviate hepatic ischemia-reperfusion injury via decreasing the Atg12-Atg5 conjugate. The expression of miR-30b, which binds the 3'-UTR of Atg12, was significantly down-regulated after HIRI. Levels of Atg12 and Atg12-Atg5 conjugate then increased and promoted autophagy, leading to apoptotic cell death. Therefore, miR-30b can inhibit autophagy to alleviate hepatic IRI by decreasing levels of Atg12-Atg5 conjugate. HIRI: Hepatic ischemia-reperfusion injury.
Figure 5 Model of miR-30b inhibition of autophagy to alleviate hepatic ischemia-reperfusion injury via decreasing the Atg12-Atg5 conjugate. The expression of miR-30b, which binds the 3'-UTR of Atg12, was significantly down-regulated after HIRI. Levels of Atg12 and Atg12-Atg5 conjugate then increased and promoted autophagy, leading to apoptotic cell death. Therefore, miR-30b can inhibit autophagy to alleviate hepatic IRI by decreasing levels of Atg12-Atg5 conjugate. HIRI: Hepatic ischemia-reperfusion injury.CC-BY-NC · From the PDF
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Metadata

Authors
Li, S.-P.
Venue
World Journal of Gastroenterology
Publisher
Baishideng Publishing Group Inc.
Published
2016
Volume
22
Issue
18
Pages
4501
ISSN
1007-9327
DOI
10.3748/wjg.v22.i18.4501
Cited by
37
References
1
Language
en

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