Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-101 inhibits autophagy to alleviate liver ischemia/reperfusion injury via regulating the mTOR signaling pathway

doi: 10.3892/ijmm.2019.4077

Figure Lengend Snippet: Alterations of miR-101 expression and autophagy signaling in the liver IR injury mouse model. (A) Relative expression levels of miR-101 and mTOR mRNA in mouse liver at each reperfusion time point were detected using reverse transcription-quantitative polymerase chain reaction. * P<0.05 and *** P<0.001 vs. miR-101 sham. # P<0.05 and ### P<0.001 vs. mTOR sham. (B) Comparison of serum ALT and AST levels at different time points. *** P<0.001 vs. ALT sham. ### P<0.001 vs. AST sham. (C) Western blots exhibiting the levels of mTOR, p-mTOR, caspase-3, p62 and LC3II proteins in the liver at different time points post-reperfusion. GAPDH was used as the internal control. * P<0.05, ** P<0.01 and *** P<0.001 vs. the sham treated group. miR, microRNA; mTOR, mechanistic target of rapamycin; IR, ischemia/reperfusion; ALT, alanine aminotransferase; AST, aspartate aminotransferase; p-, phosphorylated; LC3II, microtubule-associated protein 1 light II.

Article Snippet: Antibodies against mechanistic target of rapamycin (mTOR; cat. no. 2972), phosphorylated (p-)mTOR (cat. no. 2971), caspase-3 (cat. no. 9662), sequestosome 1/p62 (cat. no. 16177), microtubule-associated protein 1 light II (LC3II; cat. no. 3868), proliferating cell nuclear antigen (PCNA; cat. no. 13110) and GAPDH (cat. no. 5174), and the horseradish peroxidase (HRP)-conjugated anti-rabbit (cat. no. 7074) and anti-mouse (cat. no. 7076) secondary antibodies were all purchased from Cell Signaling Technology, Inc., (Danvers, MA, USA).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Comparison, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-101 inhibits autophagy to alleviate liver ischemia/reperfusion injury via regulating the mTOR signaling pathway

doi: 10.3892/ijmm.2019.4077

Figure Lengend Snippet: miR-101 ameliorates liver IR injury by inhibiting apoptosis. (A) Relative expression levels of miR-101 and mTOR mRNA in response to miR-101 agomir/antagomir were detected using reverse transcription-quantitative polymerase chain reaction. ** P<0.01 and *** P<0.001 vs. miR-101 miR-NC group. ## P<0.01 and ### P<0.001 vs. mTOR miR-NC group. (B) Representative images of haemotoxylin and eosin-stained liver sections presenting histopathological changes (×200 magnification; scale bars=50 µ m) following miR-101 agomir or antagomir injection. *** P<0.001 vs. miR-NC group. (C) Western blots presenting mTOR, caspase-3 and LC3II levels in the liver. ** P<0.01 and *** P<0.001 vs. the miR-NC group. (D) Representative images of TUNEL stained apoptotic nuclei (red) with DAPI counterstaining (×200 magnification; scale bars=50 µ m), and comparison of the percentage of apoptotic cells analyzed using Image J software. * P<0.05 and ** P<0.01 with comparisons shown by lines. miR, microRNA; mTOR, mechanistic target of rapamycin; NC, negative control; IR, ischemia/reperfusion; LC3II, microtubule-associated protein 1 light II; TUNEL, terminal uridine nick-end labeling; DAPI, 3,3′-diaminobenzidine.

Article Snippet: Antibodies against mechanistic target of rapamycin (mTOR; cat. no. 2972), phosphorylated (p-)mTOR (cat. no. 2971), caspase-3 (cat. no. 9662), sequestosome 1/p62 (cat. no. 16177), microtubule-associated protein 1 light II (LC3II; cat. no. 3868), proliferating cell nuclear antigen (PCNA; cat. no. 13110) and GAPDH (cat. no. 5174), and the horseradish peroxidase (HRP)-conjugated anti-rabbit (cat. no. 7074) and anti-mouse (cat. no. 7076) secondary antibodies were all purchased from Cell Signaling Technology, Inc., (Danvers, MA, USA).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Staining, Injection, Western Blot, TUNEL Assay, Comparison, Software, Negative Control, End Labeling

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-101 inhibits autophagy to alleviate liver ischemia/reperfusion injury via regulating the mTOR signaling pathway

doi: 10.3892/ijmm.2019.4077

Figure Lengend Snippet: miR-101 inhibits autophagy and weakens IR injury by activating the mTOR pathway in vitro . (A) Relative expression levels of miR-101 and mTOR mRNA in response to miR-101 mimetics/inhibitors. ** P<0.01 and *** P<0.001 vs. the miR-101 NC group. ## P<0.01 and ### P<0.001 vs. the mTOR NC group. (B) Western blots presenting mTOR, caspase-3 and LC3II levels in AML12 cells. * P<0.05, ** P<0.01 and *** P<0.001 vs. NC group. (C) Representative confocal images of immunofluorescent GFP-RFP-LC3 expression in AML12 cells. Yellow dots indicate the autophagosomes with GFP and RFP merging, and the red dots represent the autolysosomes with degraded GFP due to the acidic environment. Scale bars=5 µ m. * P<0.05, ** P<0.01 and *** P<0.001 with comparisons shown by lines. (D) Representative immunofluorescence images presenting the expression of p62 in AML12 at ×400 magnification. Scale bars=20 µ m. miR, microRNA; mTOR, mechanistic target of rapamycin; NC, negative control; LC3II, microtubule-associated protein 1 light II; IR, ischemia/reperfusion; GFP, green fluorescence protein; RFP, red fluorescence protein; DAPI, 3,3′-diaminobenzidine.

Article Snippet: Antibodies against mechanistic target of rapamycin (mTOR; cat. no. 2972), phosphorylated (p-)mTOR (cat. no. 2971), caspase-3 (cat. no. 9662), sequestosome 1/p62 (cat. no. 16177), microtubule-associated protein 1 light II (LC3II; cat. no. 3868), proliferating cell nuclear antigen (PCNA; cat. no. 13110) and GAPDH (cat. no. 5174), and the horseradish peroxidase (HRP)-conjugated anti-rabbit (cat. no. 7074) and anti-mouse (cat. no. 7076) secondary antibodies were all purchased from Cell Signaling Technology, Inc., (Danvers, MA, USA).

Techniques: In Vitro, Expressing, Western Blot, Immunofluorescence, Negative Control, Fluorescence

Journal: International Journal of Molecular Medicine

Article Title: MicroRNA-101 inhibits autophagy to alleviate liver ischemia/reperfusion injury via regulating the mTOR signaling pathway

doi: 10.3892/ijmm.2019.4077

Figure Lengend Snippet: Inhibition of miR-101 and mTOR expression aggravates liver IR injury. (A) Western blots presenting mTOR, p-mTOR, caspase-3, p62 and LC3II levels in AML12 cells. (B) Column graph presenting the percentage of surviving cells in each treatment group. ** P<0.01 and *** P<0.001 vs. the IR group. miR, microRNA; mTOR, mechanistic target of rapamycin; IR, ischemia/reperfusion; p-, phosphorylated; LC3II, microtubule-associated protein 1 light II; Rapa, rapamycin.

Article Snippet: Antibodies against mechanistic target of rapamycin (mTOR; cat. no. 2972), phosphorylated (p-)mTOR (cat. no. 2971), caspase-3 (cat. no. 9662), sequestosome 1/p62 (cat. no. 16177), microtubule-associated protein 1 light II (LC3II; cat. no. 3868), proliferating cell nuclear antigen (PCNA; cat. no. 13110) and GAPDH (cat. no. 5174), and the horseradish peroxidase (HRP)-conjugated anti-rabbit (cat. no. 7074) and anti-mouse (cat. no. 7076) secondary antibodies were all purchased from Cell Signaling Technology, Inc., (Danvers, MA, USA).

Techniques: Inhibition, Expressing, Western Blot