Journal: Oncology Reports

Article Title: 3-Bromopyruvate sensitizes human breast cancer cells to TRAIL-induced apoptosis via the phosphorylated AMPK-mediated upregulation of DR5

doi: 10.3892/or.2018.6644

Figure Lengend Snippet: 3-BP induces AMPK phosphorylation and induces cell death. (A) The levels of CHOP, GRP78, AMPK-α and p-AMPK were detected in breast cancer MCF-7 and MDA-MB-231 cells treated with 3-BP (0, 40, 80 and 160 µmol/l) for 24 h via western blot analysis. (B) The expression levels of Bax in MCF-7 cells and caspase-3 protein in MDA-MB-231 cells were detected via western blotting in cells treated with 80 µmol/l 3-BP and 200 ng/ml TRAIL. 3-BP, 3-bomopyruvate.

Article Snippet: The rabbit polyclonal antibodies against AMPK-α1 (1:500 dilution; cat. no. 10929-2-AP), Bax (1:5,000 dilution; cat. no. 50599-2-Ig) and Bcl-2 (1:1,000 dilution; cat. no. 12789-1-AP) were supplied by ProteinTech Group, Inc. (Chicago, IL, USA).

Techniques: Phospho-proteomics, Western Blot, Expressing

Journal: Oncology Reports

Article Title: 3-Bromopyruvate sensitizes human breast cancer cells to TRAIL-induced apoptosis via the phosphorylated AMPK-mediated upregulation of DR5

doi: 10.3892/or.2018.6644

Figure Lengend Snippet: The AMPK inhibitor Compound C attenuates the effects of 3-BP on breast cancer cells. MCF-7 and MDA-MB-231 cells were treated with 1 µmol/l Compound C (Com C), 80 µmol/l 3-BP, and 200 ng/ml TRAIL, or both 3-BP and TRAIL, as indicated. (A) Cell viability was determined using an MTT assay. (B) Cell morphology was examined via light microscopy and apoptosis rate was determined using the PI staining method and flow cytometry. Data are expressed as the mean ± standard error of the mean (n=3). TRAIL, tumor necrosis factor-related apoptosis-inducing ligand; 3-BP, 3-bomopyruvate.

Article Snippet: The rabbit polyclonal antibodies against AMPK-α1 (1:500 dilution; cat. no. 10929-2-AP), Bax (1:5,000 dilution; cat. no. 50599-2-Ig) and Bcl-2 (1:1,000 dilution; cat. no. 12789-1-AP) were supplied by ProteinTech Group, Inc. (Chicago, IL, USA).

Techniques: MTT Assay, Light Microscopy, Staining, Flow Cytometry

Journal: Oncology Reports

Article Title: 3-Bromopyruvate sensitizes human breast cancer cells to TRAIL-induced apoptosis via the phosphorylated AMPK-mediated upregulation of DR5

doi: 10.3892/or.2018.6644

Figure Lengend Snippet: AMPK induces ER stress and sensitizes breast cancer cells to TRAIL in response to treatment with 3-BP. (A) Cells treated with medium (Control), 1 µmol/l Compound C (Com C) or Compound C combined with 80 µmol/l 3-BP for 24 h were investigated via flow cytometry. (B) MCF-7 and MDA-MB-231 cells pre-treated with 1 µmol/l Compound C for 1 h were subsequently treated with 0, 40, 80 or 160 µmol/l 3-BP for 24 h. The expression levels of AMPK, GRP78, CHOP and DR5 were investigated with western blotting. (C) Cells pre-treated with or without 1 µmol/l Compound C for 1 h, were treated with medium, Compound C, 80 µmol/l 3-BP, 200 ng/ml TRAIL or both 3-BP and TRAIL, as indicated, for 24 h. The expression levels of Bax and Bcl-2 were determined in MCF-7 cells and caspase-3 was investigated in the MDA-MB-231 cells by western blotting. β-actin served as loading control. TRAIL, tumor necrosis factor-related apoptosis-inducing ligand; 3-BP, 3-bomopyruvate; DR5, death receptor 5.

Article Snippet: The rabbit polyclonal antibodies against AMPK-α1 (1:500 dilution; cat. no. 10929-2-AP), Bax (1:5,000 dilution; cat. no. 50599-2-Ig) and Bcl-2 (1:1,000 dilution; cat. no. 12789-1-AP) were supplied by ProteinTech Group, Inc. (Chicago, IL, USA).

Techniques: Control, Flow Cytometry, Expressing, Western Blot

Journal: Heliyon

Article Title: FTO-mediated autophagy inhibition promotes non-small cell lung cancer progression by reducing the stability of SESN2 mRNA

doi: 10.1016/j.heliyon.2024.e27571

Figure Lengend Snippet: Primary and secondary antibodies.

Article Snippet: Rabbit anti-AMPK alpha antibody , 1:3000 , Proteintech, USA, 10929-2-AP.

Techniques:

Journal: Heliyon

Article Title: FTO-mediated autophagy inhibition promotes non-small cell lung cancer progression by reducing the stability of SESN2 mRNA

doi: 10.1016/j.heliyon.2024.e27571

Figure Lengend Snippet: FTO-regulated SESN2 expression affects the AMPK-mTOR signaling pathway. (A) Representative immunofluorescence images of H460 and H1975 cells overexpressing SESN2 along with FTO overexpression. Immunofluorescence demonstrated the fluorescence intensity of p62 in these cells (n = 3, one-way ANOVA). Scale bar: 100 μm. (B) Western blotting to detect p -AMPKα, AMPKα, p -mTOR, mTOR expression in H460 and H1975 cells overexpressing SESN2 along with FTO overexpression (n = 3, one-way ANOVA). Data are presented as means ± SD of three independent experiments. * P < 0.05, ** P < 0.01. Uncropped versions of B were added to Supplemental Material.

Article Snippet: Rabbit anti-AMPK alpha antibody , 1:3000 , Proteintech, USA, 10929-2-AP.

Techniques: Expressing, Immunofluorescence, Over Expression, Fluorescence, Western Blot

Journal: Heliyon

Article Title: FTO-mediated autophagy inhibition promotes non-small cell lung cancer progression by reducing the stability of SESN2 mRNA

doi: 10.1016/j.heliyon.2024.e27571

Figure Lengend Snippet: Schematic diagram of the proposed mechanism by which FTO deletion-mediated autophagy inhibits the progression of NSCLC. FTO deficiency promoted the interaction between IGF2BP1 and SESN2 mRNA, which enhanced SESN2 mRNA stability and its protein expression, thereby promoting autophagy and mitigating the malignant progression of NSCLC. Besides, loss of FTO upregulated SESN2 expression, which in turn induced autophagy through increased phosphorylation of AMPKα and negative regulation of mTOR.

Article Snippet: Rabbit anti-AMPK alpha antibody , 1:3000 , Proteintech, USA, 10929-2-AP.

Techniques: Expressing, Phospho-proteomics

Journal: British Journal of Cancer

Article Title: Prognostic significance of AMP-activated protein kinase expression and modifying effect of MAPK3/1 in colorectal cancer

doi: 10.1038/sj.bjc.6605846

Figure Lengend Snippet: P hosphorylated AMPK and p-MAPK3/1 expression in colorectal cancer. ( A ) Positive for p-AMPK cytoplasmic expression (arrowheads). ( B ) Negative for p-AMPK expression (white arrowheads). ( C ) Positive for p-MAPK3/1 nuclear expression (white arrows). ( D ) Negative for p-MAPK3/1 expression (block arrow). Stromal cells serve as an internal positive control for p-MAPK3/1 expression (arrow).

Article Snippet: Primary antibody against p-AMPK (rabbit monoclonal anti-phospho-AMPK α (Thr172) (40H9), 1 : 100 dilution; Cell Signaling Technology, Boston, MA, USA) was applied ( Ji et al , 2007 ; Contreras et al , 2008 ; Hadad et al , 2009 ; Vazquez-Martin et al , 2009 ; Zheng et al , 2009 ), and the slides were maintained at 4 °C for overnight, followed by rabbit secondary antibody (Vector Laboratories) (60 min), an avidin–biotin complex conjugate (Vector Laboratories) (60 min), diaminobenzidine (5 min) and methyl-green counterstain.

Techniques: Expressing, Blocking Assay, Positive Control

Journal: British Journal of Cancer

Article Title: Prognostic significance of AMP-activated protein kinase expression and modifying effect of MAPK3/1 in colorectal cancer

doi: 10.1038/sj.bjc.6605846

Figure Lengend Snippet: p-AMPK expression in colorectal cancer, and clinical, pathologic and molecular features

Article Snippet: Primary antibody against p-AMPK (rabbit monoclonal anti-phospho-AMPK α (Thr172) (40H9), 1 : 100 dilution; Cell Signaling Technology, Boston, MA, USA) was applied ( Ji et al , 2007 ; Contreras et al , 2008 ; Hadad et al , 2009 ; Vazquez-Martin et al , 2009 ; Zheng et al , 2009 ), and the slides were maintained at 4 °C for overnight, followed by rabbit secondary antibody (Vector Laboratories) (60 min), an avidin–biotin complex conjugate (Vector Laboratories) (60 min), diaminobenzidine (5 min) and methyl-green counterstain.

Techniques: Expressing, Significance Assay, Methylation, Mutagenesis

Journal: British Journal of Cancer

Article Title: Prognostic significance of AMP-activated protein kinase expression and modifying effect of MAPK3/1 in colorectal cancer

doi: 10.1038/sj.bjc.6605846

Figure Lengend Snippet: Kaplan–Meier curves for colorectal cancer-specific survival. ( A ) p-AMPK status and survival of colorectal cancer patients. The left panel includes all eligible cases, the middle panel includes p-MAPK3/1-positive cases, and the right panel includes p-MAPK3/1-negative cases. ( B ) p-MAPK3/1 status and survival of colorectal cancer patients. The left panel includes all eligible cases, the middle panel includes p-AMPK-positive cases, and the right panel includes p-AMPK-negative cases.

Article Snippet: Primary antibody against p-AMPK (rabbit monoclonal anti-phospho-AMPK α (Thr172) (40H9), 1 : 100 dilution; Cell Signaling Technology, Boston, MA, USA) was applied ( Ji et al , 2007 ; Contreras et al , 2008 ; Hadad et al , 2009 ; Vazquez-Martin et al , 2009 ; Zheng et al , 2009 ), and the slides were maintained at 4 °C for overnight, followed by rabbit secondary antibody (Vector Laboratories) (60 min), an avidin–biotin complex conjugate (Vector Laboratories) (60 min), diaminobenzidine (5 min) and methyl-green counterstain.

Techniques:

Journal: British Journal of Cancer

Article Title: Prognostic significance of AMP-activated protein kinase expression and modifying effect of MAPK3/1 in colorectal cancer

doi: 10.1038/sj.bjc.6605846

Figure Lengend Snippet: p-AMPK status in colorectal cancer and patient mortality

Article Snippet: Primary antibody against p-AMPK (rabbit monoclonal anti-phospho-AMPK α (Thr172) (40H9), 1 : 100 dilution; Cell Signaling Technology, Boston, MA, USA) was applied ( Ji et al , 2007 ; Contreras et al , 2008 ; Hadad et al , 2009 ; Vazquez-Martin et al , 2009 ; Zheng et al , 2009 ), and the slides were maintained at 4 °C for overnight, followed by rabbit secondary antibody (Vector Laboratories) (60 min), an avidin–biotin complex conjugate (Vector Laboratories) (60 min), diaminobenzidine (5 min) and methyl-green counterstain.

Techniques:

Journal: British Journal of Cancer

Article Title: Prognostic significance of AMP-activated protein kinase expression and modifying effect of MAPK3/1 in colorectal cancer

doi: 10.1038/sj.bjc.6605846

Figure Lengend Snippet: p-AMPK status and patient mortality in strata of p-MAPK3/1 status (upper rows) and p-MAPK3/1 status and patient mortality in strata of p-AMPK status (lower rows)

Article Snippet: Primary antibody against p-AMPK (rabbit monoclonal anti-phospho-AMPK α (Thr172) (40H9), 1 : 100 dilution; Cell Signaling Technology, Boston, MA, USA) was applied ( Ji et al , 2007 ; Contreras et al , 2008 ; Hadad et al , 2009 ; Vazquez-Martin et al , 2009 ; Zheng et al , 2009 ), and the slides were maintained at 4 °C for overnight, followed by rabbit secondary antibody (Vector Laboratories) (60 min), an avidin–biotin complex conjugate (Vector Laboratories) (60 min), diaminobenzidine (5 min) and methyl-green counterstain.

Techniques:

Journal: Redox Biology

Article Title: Adiponectin/AdiopR1 signaling prevents mitochondrial dysfunction and oxidative injury after traumatic brain injury in a SIRT3 dependent manner

doi: 10.1016/j.redox.2022.102390

Figure Lengend Snippet: Transcription and expressions of SIRT3 were regulated by AdipoR1/AMPK/PGC-1α signaling pathway in TBI models. (A–D) Western blot and statistical analysis of the phosphorylation of AMPK and expressions of SIRT3 and PGC-1α. (E) The transcription results of SIRT3 in AdipoR1 flox/flox and AdipoR1 CKO mice. **p < 0.01 and ***p < 0.001 vs sham group in each strain of mice, # p < 0.05 vs TBI group in each strain of mice, & p < 0.05, && p < 0.01, ns: no statistical significance. (F–I) Effects of AMPK phosphorylation inhibitor and AdipoRon treatment on AMPK phosphorylation and expressions of SIRT3 and PGC-1α after TBI. (J) Mechanism involved in protective effects of activated APN/AdipoR1 signaling after TBI. Data are presented as mean ± SD for n = 6. **p < 0.01 vs sham group, # p < 0.05 vs TBI group, $ p < 0.05 vs TBI + AdipoRon group.

Article Snippet: The membranes were immersed in 5% non-fat milk for 2 h and incubated at 4 °C overnight in the presence of primary rabbit polyclonal antibodies against Bcl-2 (12,789, Proteintech), Bax (50,599, Proteintech), AMPK (66,536, Proteintech), p-AMPK (2537, CST), PGC-1α (66,369, Proteintech), SIRT3 (10,099, Proteintech), PRDX3 (ab264354, Abcam), acetylated-Lysine (9441, CST), VDAC (55,259, Proteintech), and β-actin (AC026, ABclonal).

Techniques: Western Blot, Phospho-proteomics