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: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

Article Title: The Cardioprotective Effect of Metformin in Doxorubicin-Induced Cardiotoxicity: The Role of Autophagy

doi: 10.3390/molecules23051184

Figure Lengend Snippet: The effect of doxorubicin and metformin on the expression level ( n = 7–8) of pAMPK/AMPK ( A ), Beclin-1 ( B ), LC3B-II ( C ), and p62 ( D ) after 2-week treatments. The expression level of proteins in left ventricular tissues was evaluated using Western blot analysis. Results are provided as average magnitude of each value within a group of animals ± SEM. The significance of differences among groups was evaluated with one-way analysis of variance (ANOVA) followed by the Tukey comparison test. p values of 0.05 or less were considered significant in each graph; * Significant difference the control group vs. DOX group; ** Significant difference DOX group vs. DOX+MET group; # Significant difference DOX group vs. MET group; † Significant difference the control group vs. MET group.

Article Snippet: Membranes were incubated overnight at 4 °C with primary antibodies directed against AMPK, p-AMPK, Beclin-1, LC3-II, p62 (Cell Signaling Technology, MA, USA).

Techniques: Expressing, Western Blot, Comparison, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Huayu Tongluo Recipe Attenuates Renal Oxidative Stress and Inflammation through the Activation of AMPK/Nrf2 Signaling Pathway in Streptozotocin- (STZ-) Induced Diabetic Rats

doi: 10.1155/2021/5873007

Figure Lengend Snippet: Effects of HTR on AMPK/Nrf2 signaling pathway. (a) Representative western blot is shown for total AMPK, pAMPK, total Nrf2, nuclear Nrf2, HO-1, NQO1, β -actin, and histone H3. Quantitative analyses are shown for (b) pAMPK/total AMPK, (c) total Nrf2/ β -actin, (d) nuclear Nrf2/H3, (e) HO-1/ β -actin, and (f) NQO1/ β -actin. (g) The mRNA relative expression of HO-1 in serum detected by qRT-PCR. (h) The mRNA relative expression of NQO1 in serum detected by qRT-PCR. (i) The protein expression detected by immunofluorescence. Data were presented as mean ± SD ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs. control; # P < 0.05, ## P < 0.01 vs. DN.

Article Snippet: The primary antibodies used in this study are as follows: AMPK α (1 : 800, Cell Signaling Technology, USA, 5831), p-AMPK α (1 : 1000, 2535), Nrf2 (1 : 1000, Abcam, USA, 89443), Nox4 (1 : 1000, 133303), TGF- β 1 (1 : 600, 215715), HO-1 (1 : 1000, Santa Cruz, USA, 390991), NQO1 (1 : 1000, 376023), and NF κ B (1 : 600, Servicebio, China, 11997).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Immunofluorescence, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Huayu Tongluo Recipe Attenuates Renal Oxidative Stress and Inflammation through the Activation of AMPK/Nrf2 Signaling Pathway in Streptozotocin- (STZ-) Induced Diabetic Rats

doi: 10.1155/2021/5873007

Figure Lengend Snippet: Schematic overview of HTR's effect on renal fibrosis in STZ-induced diabetic rats. ECM accumulation, the characteristic of fibrosis, is a main pathological change in DN. As we all know, oxidative stress and inflammation, which promote each other, are the critical causes of TGF- β 1-induced excessive synthesis and deposition of ECM in DN. Interestingly, AMPK acts as the upstream of Nrf2. The activation of AMPK/Nrf2 pathway can inhibit the Nox4-induced oxidative stress and attenuate NF κ B-induced inflammation. HTR enhances the activity of AMPK/Nrf2 pathway and suppresses oxidative stress and inflammation, thereby decreasing the ECM deposition and delaying the progression of fibrosis in DN (⟶: activated; ⊣: inhibited).

Article Snippet: The primary antibodies used in this study are as follows: AMPK α (1 : 800, Cell Signaling Technology, USA, 5831), p-AMPK α (1 : 1000, 2535), Nrf2 (1 : 1000, Abcam, USA, 89443), Nox4 (1 : 1000, 133303), TGF- β 1 (1 : 600, 215715), HO-1 (1 : 1000, Santa Cruz, USA, 390991), NQO1 (1 : 1000, 376023), and NF κ B (1 : 600, Servicebio, China, 11997).

Techniques: Activation Assay, Activity Assay