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: Scientific Reports

Article Title: AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N 6 -methyladenosine

doi: 10.1038/srep41606

Figure Lengend Snippet: ( A,B ) The triglyceride content in SOL was much higher than EDL ( A ) while the p-AMPKα2 level in SOL was lower than EDL ( B ). ( C,D ) The EDL of OB mice contained more triglyceride ( C ) but a lower p-AMPKα2 level compared to WT mice ( D ). ( E,F ) The triglyceride content ( E ) and the p-AMPKα2 levels ( F ) in SOL of OB mice ( F ). The results are presented as the mean ± SEM. n = 4–6. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: siRNA against FTO or AMPKα2 and a negative control (siCTRL) were designed and produced by RiboBio Company (Guangzhou, China).

Techniques:

Journal: Scientific Reports

Article Title: AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N 6 -methyladenosine

doi: 10.1038/srep41606

Figure Lengend Snippet: ( A ) The p-AMPKα2 level was significantly inhibited by Compound C. ( B,C ) ORO staining ( B ) and triglyceride content ( C ) in C2C12 cells treated withor without Compound C. ( D ) AICAR upregulates p-AMPKα2 expression in C2C12 cells. ( E,F ) ORO staining ( E ) and triglyceride content ( F ) in C2C12 cells treated withor without AICAR. Data are presented as the mean ± SEM. n = 3. * p < 0.05, ** p < 0.01.

Article Snippet: siRNA against FTO or AMPKα2 and a negative control (siCTRL) were designed and produced by RiboBio Company (Guangzhou, China).

Techniques: Staining, Expressing

Journal: Scientific Reports

Article Title: AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N 6 -methyladenosine

doi: 10.1038/srep41606

Figure Lengend Snippet: ( A ) The mRNA levels of AMPKα2 in C2C12 cells treated with AMPKα2 siRNA (siAMPKα2). ( B–D ) m 6 A methylation in C2C12 cells treated with siAMPKα2 ( B ), CC ( C ) or AICAR ( D ). ( E ) The m 6 A levelsin C2C12 cells treated with CL, AICAR or their combination. ( F,G ) ORO staining ( F ) and TG content ( G ) in C2C12 cells treated with CL, AICAR or their combination. ( H,I ) The mRNA and protein levels of the adipogenic and fatty acid lipolysis related genesin C2C12 cells treated with CL, AICAR or their combination. CC, Compound C; CL, cycloleucine. The results are presented as the mean ± SEM. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bars: 100 μm.

Article Snippet: siRNA against FTO or AMPKα2 and a negative control (siCTRL) were designed and produced by RiboBio Company (Guangzhou, China).

Techniques: Methylation, Staining

Journal: Scientific Reports

Article Title: AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N 6 -methyladenosine

doi: 10.1038/srep41606

Figure Lengend Snippet: ( A,B ) The protein levels of AMPKα2, FTO, METTL3 and METTL14 in C2C12 cells after knockdown of AMPKα2. The relative expression levels of AMPKα2, METTL3, METTL14 and FTO were normalized to GAPDH. ( C,D ) The protein levels of AMPKα2, p-AMPKα2, FTO, METTL3 and METTL14 in C2C12 cells after AICAR treatment. The relative protein levels of p-AMPKα2, METTL3, METTL14 and FTO were normalized to GAPDH in C2C12 cells after AICAR treatment. ( E ) The m 6 A methylation levels after treatment with siAMPKα2, siFTO or their combination. ( F ) The triglyceride content after treatment with siAMPKα2, siFTO or their combination. ( G ) A model depicting the role of AMPK in the regulation of lipid accumulation and m 6 A methylation through FTO-dependent demethylation in skeletal muscle cells. CC, Compound C. Data are presented as the mean ± SEM. n = 3. * p < 0.05, ** p < 0.01.

Article Snippet: siRNA against FTO or AMPKα2 and a negative control (siCTRL) were designed and produced by RiboBio Company (Guangzhou, China).

Techniques: Knockdown, Expressing, Methylation

Journal: Scientific Reports

Article Title: AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N 6 -methyladenosine

doi: 10.1038/srep41606

Figure Lengend Snippet: Primers used for real-time PCR.

Article Snippet: siRNA against FTO or AMPKα2 and a negative control (siCTRL) were designed and produced by RiboBio Company (Guangzhou, China).

Techniques:

Journal: Molecular cell

Article Title: Phosphorylation of EZH2 by AMPK Suppresses PRC2 Methyltransferase Activity and Oncogenic Function

doi: 10.1016/j.molcel.2017.12.024

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: AMPK (A1/B1/G2) Kinase , Promega , Cat # V4012.

Techniques: Virus, Recombinant, Purification, Sample Prep, Sequencing, Software

Journal: Molecular Vision

Article Title: Aqueous cytokine and growth factor levels do not reliably reflect those levels found in the vitreous

doi:

Figure Lengend Snippet: List of antibodies and manufactures used in this study.

Article Snippet: AMPK α1 S485 , Hypoxia/Ischemia , Assay Designs , Plymouth Meeting, PA.

Techniques: