Journal: The Journal of international medical research

Article Title: The therapeutic potential of irisin in alleviating acute lung injury via inflammation and ferroptosis modulation.

doi: 10.1177/03000605251340338

Figure Lengend Snippet: Figure 6. Western blotting analysis on the effects of irisin on the expression level of ferroptosis-associated proteins. (a) Representative western blotting images for the expression of ACSL4, COX-2, GPX4, p-AMPK, and t-AMPK in lung tissues. GAPDH was selected as the loading control protein. (b) Quantification analysis of the related bands of ACSL4, COX-2, GPX4, p-AMPK, and t-AMPK in lung tissues. Lung tissues were harvested on day 3 post-CLP. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. n ¼ 6 per group. Data are presented as means SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Subsequently, the membranes were incubated with rabbit anti-AMPK (MCE, Cat# HY-P80541), anti-p-AMPK (MCE, HYP80452), GPX4 (MCE, HY-P80450), ACSL4 (Santa Cruz Biotechnology, Dallas, Texas, USA; Cat# sc-365230), COX-2 (Abcam, Cambridge, United Kingdom; Cat# ab283574), and GAPDH (MCE, HY-P80137) antibodies at 4 C overnight.

Techniques: Western Blot, Expressing, Control

Journal: Journal of Agricultural and Food Chemistry

Article Title: Natural Dietary Flavonoid Apigenin Mitigates Ulcerative Colitis via Modulating the AMPK/NF-κB/NLRP3 Signaling Axis

doi: 10.1021/acs.jafc.5c15405

Figure Lengend Snippet: Api alleviates colitis by coordinately modulating AMPK and NLRP3. (A-B) ELISA quantification of TNF-α, IL-6 (A), and IL-1β (B) in BMDM supernatants following LPS + nigericin stimulation with or without Api, the AMPK agonist AICAR, or the AMPK inhibitor BAY-3827 ( n = 4). (C) Effect of AMPKα1 knockdown on the mRNA expression of Prkaa1 , TNF-α , and Nlrp3 in BMDMs subjected to LPS + nigericin stimulation with or without Api treatment ( n = 6). (D) SPR sensorgram confirming the direct interaction between Api and the recombinant AMPK α1/β2/γ1 complex. (E) Molecular-docking models depicting Api bound to multiple reported AMPK allosteric or activator-related pockets, including AMPKα1 (PDB: 6C9F), AMPKα1β1γ1 (PDB: 6C9F), AMPKα1β1γ1 (PDB: 4QFR), and AMPKα2β1γ1 (PDB: 4CFF).

Article Snippet: Commercially available recombinant human NLRP3 protein (CSB-EP822275HU7) and AMPK α1/β2/γ1 heterotrimer protein (HY- P76143 , MCE) were immobilized (∼3,000 RU) on a CM5 Chip (GE Healthcare, Piscataway, NJ, USA) according to a standard amine coupling procedure.

Techniques: Enzyme-linked Immunosorbent Assay, Knockdown, Expressing, Recombinant

Journal: Viruses

Article Title: The Function behind the Relation between Lipid Metabolism and Vimentin on H9N2 Subtype AIV Replication.

doi: 10.3390/v14081814

Figure Lengend Snippet: Figure 4. The cellular AMPK expression and phosphorylation level. AMPK protein expressions and phosphorylation in HeLa and KO HeLa cells with or without viral infection were detected with Western blotting. (A) The expression and phosphorylation of AMPK protein on KO HeLa cells compared with HeLa cells. (B) The expression and phosphorylation of AMPK protein on HeLa cells after viral infection. (C) The expression and phosphorylation of AMPK protein on KO HeLa cells after viral infection.

Article Snippet: GAPDH monoclonal antibody was purchased from Enogene Biologicals (Nanjing, China). β-actin monoclonal antibody, AMPK monoclonal antibody and AMPK phosphorylated monoclonal antibody P-AMPK were purchased from BOSTER Biologicals (Nanjing, China).

Techniques: Expressing, Phospho-proteomics, Infection, Western Blot

Journal: Viruses

Article Title: The Function behind the Relation between Lipid Metabolism and Vimentin on H9N2 Subtype AIV Replication.

doi: 10.3390/v14081814

Figure Lengend Snippet: Figure 7. Model of lipid metabolism and vimentin on H9N2 subtype AIV. When vimentin was knocked down, the phosphorylation level of AMPK was decreased, resulting in the decreased level of HMGCR phosphorylation, which increased the enzyme activity, thereby increasing cholesterol. After the destruction of lipid rafts, the virus binding was inhibited, and cholesterol helped to stabilize the structure of lipid rafts, which might help the virus bind to cells.

Article Snippet: GAPDH monoclonal antibody was purchased from Enogene Biologicals (Nanjing, China). β-actin monoclonal antibody, AMPK monoclonal antibody and AMPK phosphorylated monoclonal antibody P-AMPK were purchased from BOSTER Biologicals (Nanjing, China).

Techniques: Phospho-proteomics, Activity Assay, Virus, Binding Assay

Journal: Journal of Functional Foods

Article Title: Anti-fatigue effects and mechanism of Syngnathus schlegeli peptides supplementation on exercise-fatigued mice

doi: 10.1016/j.jff.2023.105846

Figure Lengend Snippet: Fig. 4. Effects of SAP on protein expression in gastrocnemius muscle were evaluated using western blotting. (A-F) The protein levels of NRF2, p-AMPK/AMPK, PGC- 1α, SIRT1, NRF1and TFAM were normalized to GAPDH and their relative band intensities were quantified. Data were shown as the mean ± SD from six independent experiments. Different letters indicate significant differences (p < 0.05) between values in each group.

Article Snippet: Alkaline protease (200000 U/g) was obtained from Beijing Solarbio Science & Technology CO., Ltd. Antibodies used in this study include AMPK, p-AMPK, SIRT1, PGC-1α, NRF1, TFAM, GAPDH and HRP, all purchased from Boster Biological Technology Co., Ltd. (Wuhan, China) and NRF2 was obtained from ABclonal Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Western Blot