Journal: World Journal of Gastroenterology

Article Title: Antagonizing adipose tissue-derived exosome miR-103-hepatocyte phosphatase and tensin homolog pathway alleviates autophagy in non-alcoholic steatohepatitis: A trans-cellular crosstalk

doi: 10.3748/wjg.v29.i29.4528

Figure Lengend Snippet: The capacity of miR-103 in targeting phosphatase and tensin homolog gene and affecting autophagy. A: The luciferase reporter assay results verified the interaction between miR-103 and phosphatase and tensin homolog; B: Western blotting showed the differential expression of the autophagy-related protein; C: Transmission electron microscopy images of autophagosomes (red arrowhead) in the liver; D: The results of immunofluorescence staining showed liver autophagosomes in different groups. a P < 0.01 vs control; b P < 0.01 vs model; c P < 0.01 vs miR-103; d P < 0.05 vs model. PTEN: Phosphatase and tensin homolog; WT: Wild type; MUT: Mutant; DAPI: 4’,6-diamidino-2-phenylindole; mTOR: Mammalian target of rapamycin; NC: Negative control.

Article Snippet: The membranes were blocked and incubated overnight with antibodies against PTEN (9188T, CST), p-AMPK (ab32047, Abcam), p-mammalian target of rapamycin (mTOR) (CSB-PA271384, Cusabio), LC3 (12741T, CST), p62 (ab91526, Abcam), and GAPDH (ab245355, Abcam) at 4 °C.

Techniques: Luciferase, Reporter Assay, Western Blot, Quantitative Proteomics, Transmission Assay, Electron Microscopy, Immunofluorescence, Staining, Control, Mutagenesis, Negative Control

Journal: World Journal of Gastroenterology

Article Title: Antagonizing adipose tissue-derived exosome miR-103-hepatocyte phosphatase and tensin homolog pathway alleviates autophagy in non-alcoholic steatohepatitis: A trans-cellular crosstalk

doi: 10.3748/wjg.v29.i29.4528

Figure Lengend Snippet: The effect of adipose tissue-derived exosomes miR-103 on autophagy in mice. A and B: Western blotting detected the expression of the autophagy-related protein; C: The results of immunofluorescence staining to observe autophagosomes in the liver from different groups; D: Transmission electron microscopy images of autophagosomes (red arrowhead) in the liver from different groups. a P < 0.05 vs control; b P < 0.01 vs control; c P < 0.05 vs exosomes; d P < 0.01 vs exosomes. Exo: Exosomes; PTEN: Phosphatase and tensin homolog; DAPI: 4’,6-diamidino-2-phenylindole; mTOR: Mammalian target of rapamycin; NC: Negative control.

Article Snippet: The membranes were blocked and incubated overnight with antibodies against PTEN (9188T, CST), p-AMPK (ab32047, Abcam), p-mammalian target of rapamycin (mTOR) (CSB-PA271384, Cusabio), LC3 (12741T, CST), p62 (ab91526, Abcam), and GAPDH (ab245355, Abcam) at 4 °C.

Techniques: Derivative Assay, Western Blot, Expressing, Immunofluorescence, Staining, Transmission Assay, Electron Microscopy, Control, Negative Control

Journal: Molecular and Cellular Biochemistry

Article Title: Low glucose dependent decrease of apoptosis and induction of autophagy in breast cancer MCF-7 cells

doi: 10.1007/s11010-016-2711-4

Figure Lengend Snippet: Western blot ( a ) and densitometric analysis ( b ) of mTOR kinase expression in MCF-7 cells incubated in high glucose (H) and low glucose (L) DMEM for 12, 24, and 48 h. Samples containing 30 μg of protein were submitted to electrophoresis and immunoblotting. Densitometric analysis was presented as relative protein expression. The expression of β-tublin served as a control for protein loading. A representative Western blot from one of three independent experiments is presented ( a ). Mean values of densitometric analysis from three independent experiments ± SD are presented ( b )

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM), containing glucose at 4.5 mg/ml (25 mM), Dulbecco’s modified Eagle’s medium (DMEM), containing glucose at 0.5 mg/ml (2.8 mM), l -glutamine, penicillin, streptomycin, trypsin–EDTA, FBS Gold, trypsin–EDTA were provided by Gibco (San Diego, USA), passive lysis buffer by Promega (Madison, USA), BCA Protein Assay Kit by Thermo Scientific (Rockford, USA), PE Annexin V Apoptosis Detection Kit I by BD PharmingenTM (CA, USA), Sigma-Fast BCIP/NBT reagent, 4′,6-diamidino-2-phenylindole dihydrochloride—DAPI, donkey serum, acridine orange, ethidium bromide, camptothecin by Sigma (St Louis, MO, USA), monoclonal (mouse) anti-human ORP150 antibody by IBL (Gunma, Japan), medium coverquick by Hygeco (USA), polyclonal (rabbit) anti-human NF-κB2 p100/p52 antibody, monoclonal (rabbit) anti-human CHOP antibody, monoclonal (rabbit) anti-human β-tubulin antibody, alkaline phosphatase-labeled anti-rabbit immunoglobulin G were provided by Cell Signaling Technology (Boston, USA), monoclonal (mouse) anti-human mTOR antibody (Merck-Millipore, USA), alkaline phosphatase-labeled anti-mouse immunoglobulin G by Rockland (PA, USA), polyclonal (rabbit) anti-human LC3 by Abgen, donkey anti-rabbit IgG conjugated with Alexa Fluor 488 by Molecular Probes (USA).

Techniques: Western Blot, Expressing, Incubation, Electrophoresis, Control

Journal: Molecular and Cellular Biochemistry

Article Title: Low glucose dependent decrease of apoptosis and induction of autophagy in breast cancer MCF-7 cells

doi: 10.1007/s11010-016-2711-4

Figure Lengend Snippet: The effect of glucose shortage on apoptosis, autophagy, and their mechanism in breast cancer MCF-7 cell line. ORP150 oxygen-regulated protein 150, UPR unfolded protein response, NF-κB nuclear factor-κB, mTOR mammalian target of rapamycin, ATP adenosine triphosphate, CHOP CCAAT/enhancer-binding protein (C/EBP) homologous protein, P53 protein p53, P27 protein p27

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM), containing glucose at 4.5 mg/ml (25 mM), Dulbecco’s modified Eagle’s medium (DMEM), containing glucose at 0.5 mg/ml (2.8 mM), l -glutamine, penicillin, streptomycin, trypsin–EDTA, FBS Gold, trypsin–EDTA were provided by Gibco (San Diego, USA), passive lysis buffer by Promega (Madison, USA), BCA Protein Assay Kit by Thermo Scientific (Rockford, USA), PE Annexin V Apoptosis Detection Kit I by BD PharmingenTM (CA, USA), Sigma-Fast BCIP/NBT reagent, 4′,6-diamidino-2-phenylindole dihydrochloride—DAPI, donkey serum, acridine orange, ethidium bromide, camptothecin by Sigma (St Louis, MO, USA), monoclonal (mouse) anti-human ORP150 antibody by IBL (Gunma, Japan), medium coverquick by Hygeco (USA), polyclonal (rabbit) anti-human NF-κB2 p100/p52 antibody, monoclonal (rabbit) anti-human CHOP antibody, monoclonal (rabbit) anti-human β-tubulin antibody, alkaline phosphatase-labeled anti-rabbit immunoglobulin G were provided by Cell Signaling Technology (Boston, USA), monoclonal (mouse) anti-human mTOR antibody (Merck-Millipore, USA), alkaline phosphatase-labeled anti-mouse immunoglobulin G by Rockland (PA, USA), polyclonal (rabbit) anti-human LC3 by Abgen, donkey anti-rabbit IgG conjugated with Alexa Fluor 488 by Molecular Probes (USA).

Techniques: Binding Assay