Journal: Nutrients

Article Title: Adiponectin Signaling Modulates Fat Taste Responsiveness in Mice

doi: 10.3390/nu16213704

Figure Lengend Snippet: An AMPK inhibitor (Compound C) eliminates AdipoRon’s ability to enhance LA-induced calcium responses in mouse taste bud cells. The enhancement of taste cell responses to 30 μM LA caused by application of 5 μM AdipoRon was effectively eliminated by the addition of 10 μM Compound C in mouse taste bud cells from ( A ) male circumvallate (F*(2, 51.63) = 10.97, p = 0.0001; W(2, 58.43) = 8.772, p = 0.0005), ( B ) male fungiform (F(2, 105) = 3.562, p = 0.0319), ( C ) female circumvallate (F*(2, 48.23) = 11.35, p < 0.0001; W(2, 60.16) = 6.545, p = 0.0027), and ( D ) female fungiform (F*(2, 137.6) = 8.508, p = 0.0003; W(2, 103.3) = 7.086, p = 0.0013) taste cells. Data are presented as mean ± SEM. One-way ordinary ANOVA with Tukey’s test for multiple comparisons was used to determine statistical significance with equal standard deviations. Brown–Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test were introduced when standard deviations significantly differed. The letters above the bars indicate statistically significant groups within each cell type.

Article Snippet: Stock solutions of AdipoRon (MedChem Express, Monmouth Junction, NJ, USA), sulfosuccinimidyl oleate (SSO; Cayman Chemical, Ann Arbor, MI, USA), and dorsomorphin (Compound C; ApexBio, Houston, TX, USA) were made in DMSO and diluted the day of the experiment to a designated concentration.

Techniques:

Journal: Brain, behavior, and immunity

Article Title: Chemerin suppresses neuroinflammation and improves neurological recovery via CaMKK2/AMPK/Nrf2 pathway after germinal matrix hemorrhage in neonatal rats.

doi: 10.1016/j.bbi.2018.02.015

Figure Lengend Snippet: Fig. 8. Selective inhibition of ChemR23/CAMKK2/AMPK signaling in activated microglial cells abolished the effect of rh-Chemerin on inhibiting neuroinflammation at 72 h after GMH. (A) Representative images of immunofluorescence staining showing fluorescently labeled liposomes were engulfed almost exclusively in Iba1 + microglia (Arrows, red (a–c)) rather than in GFAP+ (Arrowheads, red (d)) and NeuN+ (Arrowheads, red (e)) cells at 72 h after GMH. (B) Representative images of Western blot data showing the expression of pCAMKK2, pAMPK, and Nrf2, as well as IL-1beta, IL-6, and TNF alpha either with rh-Chemerin treatment alone, rh-Chemerin + Lipo-Alpha-NETA, rh-Chemerin + Lipo-STO-609 or rh-Chemerin + Lipo-Dorsomorphin. (C) Western blot analysis of ChemR23 showed that ChemR23 level does not change with Lipo-Alpha-NETA, Lipo-STO-609 or Lipo-Dorsomorphin intervention. (D) Western blot analysis of pCAMKK2 to CAMKK2 ratio showed that pCAMKK2 increased in the rh-Chemerin treatment group and decreased in Lipo-Alpha-NETA and Lipo-STO-609 groups. However, pCAMKK2 level does not change with Lipo-Dorsomorphin intervention. (E) Western blot analysis of pAMPK to AMPK ratio showed that pAMPK increased in the rh-Chemerin treatment group and decreased in Lipo-Alpha-NETA, Lipo-STO-609 and Lipo-Dorsomorphin groups. (F) Western blot analysis of Nrf2 showed that Nrf2 increased in the rh-Chemerin treatment group and decreased in Lipo-Alpha-NETA, Lipo-STO-609 and Lipo-Dorsomorphin groups. (G-I) Western blot data showed that Lipo-Alpha-NETA, Lipo-STO-609 and Lipo-Dorsomorphin reversed the inhibitory effects of rh-Chemerin on pro-inflammatory cytokines. (All samples of GMH + rh-Chemerin in Western blot were from the same animals which were euthanized after short-term neurobehavioral tests). Data are mean ± SD. ANOVA, Dunnett. * p < 0.05 compared to sham, # p < 0.05 compared to GMH + vehicle, N = 6–8/group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Liposomes (FormuMax) that contain a lipid fluorescent dye, Fluorescein DHPE (Lipo-DHPE), Alpha-NETA (ChemR23 specific inhibitor, Lipo-Alpha-NETA, Santa Cruz Biotechnology) (Graham et al., 2014), STO-609 (CAMKK2 specific inhibitor, Lipo-STO-609, Santa Cruz Biotechnology) (Cary et al., 2013) or Dorsomorphin (AMPK specific inhibitor, Lipo-Dorsomorphin, Santa Cruz Biotechnology) (Shi et al., 2017) were prepared according to the manufacturer’s protocol.

Techniques: Inhibition, Staining, Labeling, Liposomes, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Therapeutic metformin/AMPK activation blocked lymphoma cell growth via inhibition of mTOR pathway and induction of autophagy

doi: 10.1038/cddis.2012.13

Figure Lengend Snippet: Dysregulation of AMPK/mTOR pathway in human lymphoma. Expression profile of phosphorylated form of AMPK, mTOR, as well as its downstream effectors p70S6K and 4EBP1 was assessed in human lymphoma tissue using tissue array. The melanoma tissue was served as the positive control of phosphorylated AMPK

Article Snippet: The AMPK inhibitor compound C and the autophagy inhibitor 3-methyladenine were from Merck KGaA (Darmstadt, Germany).

Techniques: Expressing, Positive Control

Journal: Cell Death & Disease

Article Title: Therapeutic metformin/AMPK activation blocked lymphoma cell growth via inhibition of mTOR pathway and induction of autophagy

doi: 10.1038/cddis.2012.13

Figure Lengend Snippet: The inhibitory effect of metformin on growth of human lymphoma cell lines. ( a ) IC 50 results obtained from MTT assay in a series of B- and T-lymphoma cell lines after metformin treatment. ( b and c ) Response curves ( b ) and AMPK/ACC expression detected by western blot ( c ) in malignant B and T cells treated with metformin. ( d ) Metformin inhibited growth of primary lymphoma cells. ( e ) The proliferation of CD34+ cells enriched from cord blood samples was not altered by metformin up to 120 mM

Article Snippet: The AMPK inhibitor compound C and the autophagy inhibitor 3-methyladenine were from Merck KGaA (Darmstadt, Germany).

Techniques: MTT Assay, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Therapeutic metformin/AMPK activation blocked lymphoma cell growth via inhibition of mTOR pathway and induction of autophagy

doi: 10.1038/cddis.2012.13

Figure Lengend Snippet: Metformin-induced mTOR inhibition in an AMPK-dependent manner. ( a ) Metformin decreased the expression of phosphorylated mTOR, as well as its effectors p70S6K and 4EBP1. ( b and c ) Specific AMPK inhibitor compound C ( b ) and AMPK siRNA ( c ) abrogated metformin-induced lymphoma cell growth inhibition and cell cycle arrest. Comparing with the negative control (Con siRNA), Jurkat cells transfected with AMPK- α siRNA (AMPK siRNA) failed to induce phosphorylated ACC and inhibit phosphorylated mTOR. * P <0.001 comparing with the metformin group. + P <0.001 comparing with the Con siRNA group. ( d ) The expression of phosphorylated AKT remained unchanged during metformin treatment in Jurkat cells. ( e ) Cell sensitivity to metformin was not affected in Jurkat cells transfected with AKT

Article Snippet: The AMPK inhibitor compound C and the autophagy inhibitor 3-methyladenine were from Merck KGaA (Darmstadt, Germany).

Techniques: Inhibition, Expressing, Negative Control, Transfection

Journal: Cell Death & Disease

Article Title: Therapeutic metformin/AMPK activation blocked lymphoma cell growth via inhibition of mTOR pathway and induction of autophagy

doi: 10.1038/cddis.2012.13

Figure Lengend Snippet: The effect of metformin in murine lymphoma xenograft models. ( a ) In both B- and T-cell lymphoma models, intraperitoneal treatment of metformin, both at the high-dose (IP MET H) and low-dose (IP MET L) group, significantly diminished xenograft tumor size. ( b ) As compared with the control group (Con), decreased proliferative index Ki-67 was identified, while TUNEL staining was negative in the metformin group (MET). Immunohistochemical study showed increased numbers of cells positive for phosphorylated AMPK, but decreased cells positive for phosphorylated mTOR in the metformin-treated tumors

Article Snippet: The AMPK inhibitor compound C and the autophagy inhibitor 3-methyladenine were from Merck KGaA (Darmstadt, Germany).

Techniques: Control, TUNEL Assay, Staining, Immunohistochemical staining

Journal: Cell Death & Disease

Article Title: Therapeutic metformin/AMPK activation blocked lymphoma cell growth via inhibition of mTOR pathway and induction of autophagy

doi: 10.1038/cddis.2012.13

Figure Lengend Snippet: Metformin enhanced lymphoma cell sensitivity to the chemotherapeutic agent and stimulate autophagy. ( a ) In Daudi and Jurkat cells, metformin increased the cytotoxicity of the chemotherapeutic agent doxorubicin. ( b ) Metformin combined with doxorubicin induced less lymphoma cell growth inhibition in the AMPK siRNA group than in the Con siRNA group, * P <0.001 comparing with the Con siRNA group. ( c ) Autophagy-related LC3-II expression was significantly elevated in cells co-treated with metformin and doxorubicin. ( d ) Autophagy inhibitor 3-methyladenine abrogated lymphoma cell growth inhibition induced by co-treatment. ( e ) Combined oral metformin with doxorubicin significantly inhibited xenograft lymphoma growth, comparing with the control group or each agent alone. ( f ) The metformin/doxorubicin combination frequently induced lymphoma cell autophagy

Article Snippet: The AMPK inhibitor compound C and the autophagy inhibitor 3-methyladenine were from Merck KGaA (Darmstadt, Germany).

Techniques: Inhibition, Expressing, Control

Journal: Cell Death & Disease

Article Title: Therapeutic metformin/AMPK activation blocked lymphoma cell growth via inhibition of mTOR pathway and induction of autophagy

doi: 10.1038/cddis.2012.13

Figure Lengend Snippet: Metformin potentiated the effect of mTOR inhibitor to induce lymphoma cell autophagy. ( a ) In Daudi and Jurkat cells, metformin enhanced the growth-inhibitory effect of the mTOR inhibitor temsirolimus. ( b ) Metformin combined with temsirolimus induced less lymphoma cell growth inhibition in the AMPK siRNA group than in the Con siRNA group, * P <0.001 comparing with the Con siRNA group. ( c ) Co-treatment resulted in LC3-II upregulation. ( d ) Lymphoma cell growth inhibition was overcome by autophagy inhibitor 3-methyladenine. ( e ) Combined oral metformin with temsirolimus significantly inhibited xenograft lymphoma growth, comparing with the control group or each agent alone. ( f ) The metformin/temsirolimus combination frequently induced lymphoma cell autophagy

Article Snippet: The AMPK inhibitor compound C and the autophagy inhibitor 3-methyladenine were from Merck KGaA (Darmstadt, Germany).

Techniques: Inhibition, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Sodium Butyrate Ameliorates Oxidative Stress-Induced Intestinal Epithelium Barrier Injury and Mitochondrial Damage through AMPK-Mitophagy Pathway

doi: 10.1155/2022/3745135

Figure Lengend Snippet: Effects of sodium butyrate (NaB) on oxidative stress, intestinal epithelium barrier, and mitophagy of porcine intestinal epithelial cells (IPEC-J2) after inhibiting mitophagy or AMPK. (a–c) Superoxide dismutase (SOD), glutathione reductase (GSH) activity, and malondialdehyde (MDA) content of IPEC-J2 treated with Mdivi-1 or Compound C (CC). (d) Cellular mitochondrial membrane potential quantification by flow cytometry. (e) Cellular reactive oxygen species (ROS) level quantification by flow cytometry. (f) Intestinal epithelial transepithelial resistance (TER) and fluorescein isothiocyanate dextran 4 kDa (FD4) permeability. (g) Protein expression and quantification of tight junction Claudin-1, Occludin, and ZO-1. ∗ indicates a significant difference compared with the control group ( P < 0.05); # indicates a significant difference compared with the H2O2 group ( P < 0.05); & indicates a significant difference compared with the NaB+H2O2 group ( P < 0.05).

Article Snippet: Trypsin (Beyotime Biotechnology, China), phosphate-buffered saline (PBS) (Bozan Biotechnology, China), penicillin-streptomycin (Solabao Biotechnology, China), fetal bovine serum (Gemini, Australia), CCK-8 kit (Beyotime Biotechnology, China), MitoSpyTM Red CMXRos (Biolegend, USA), immunofluorescence fixative (Sevier Biotechnology, China), Triton-X 100 (Sigma-Aldrich, St. Louis, MO, USA), Glycine (Sinopharm Group, China), DAPI (Beyotime Biotechnology, China), Goat Anti-Mouse IgG Dylight 594 (Earthox, USA), Goat Anti-Mouse IgG Dylight 488 (Earthox, USA), mitochondrial division inhibitor (Mdivi-1) (Selleck, USA), AMPK inhibitor (Compound C, CC) (Selleck, USA), Lipofectamine RNAiMAX, and Lipofectamine 2000 were obtained from Invitrogen (Invitrogen, USA).

Techniques: Activity Assay, Membrane, Flow Cytometry, Permeability, Expressing, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Sodium Butyrate Ameliorates Oxidative Stress-Induced Intestinal Epithelium Barrier Injury and Mitochondrial Damage through AMPK-Mitophagy Pathway

doi: 10.1155/2022/3745135

Figure Lengend Snippet: Effects of sodium butyrate (NaB) on mitophagy, oxidative stress, and intestinal epithelium barrier after interference with AMPK α . (a) Expression and quantification of mitophagy proteins PINK1, Parkin, and P62. (b–d) Superoxide dismutase (SOD), glutathione reductase (GSH) activity, and malondialdehyde (MDA) content of porcine intestinal epithelial cells (IPEC-J2). (e) Cellular reactive oxygen species (ROS) level of IPEC-J2. (f) Cellular mitochondrial membrane potential of IPEC-J2. (g) Protein expression and quantification of recombinant NLR family, pyrin domain-containing protein 3 (NLRP3) and Caspase-1. (h) Intestinal epithelial transepithelial resistance (TER) and fluorescein isothiocyanate dextran 4 kDa (FD4) permeability. ∗ indicates a significant difference compared with the control group ( P < 0.05).

Article Snippet: Trypsin (Beyotime Biotechnology, China), phosphate-buffered saline (PBS) (Bozan Biotechnology, China), penicillin-streptomycin (Solabao Biotechnology, China), fetal bovine serum (Gemini, Australia), CCK-8 kit (Beyotime Biotechnology, China), MitoSpyTM Red CMXRos (Biolegend, USA), immunofluorescence fixative (Sevier Biotechnology, China), Triton-X 100 (Sigma-Aldrich, St. Louis, MO, USA), Glycine (Sinopharm Group, China), DAPI (Beyotime Biotechnology, China), Goat Anti-Mouse IgG Dylight 594 (Earthox, USA), Goat Anti-Mouse IgG Dylight 488 (Earthox, USA), mitochondrial division inhibitor (Mdivi-1) (Selleck, USA), AMPK inhibitor (Compound C, CC) (Selleck, USA), Lipofectamine RNAiMAX, and Lipofectamine 2000 were obtained from Invitrogen (Invitrogen, USA).

Techniques: Expressing, Activity Assay, Membrane, Recombinant, Permeability, Control