Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Standardized Kaempferia parviflora Wall. ex Baker (Zingiberaceae) Extract Inhibits Fat Accumulation and Muscle Atrophy in ob/ob Mice

doi: 10.1155/2018/8161042

Figure Lengend Snippet: Effect of KPE on adiposity in epididymal fat. (a) Relative protein levels of p-AMPK, AMPK, p-ACC, and ACC. (b) Relative protein levels of PPAR γ , C/EBP α , and SREBP-1c. (c) Relative mRNA levels of FAS, ACC1, LPL, and HMGR. (d) Relative protein levels of PGC-1 α , UCP2, and UCP3. ## p < 0.01 compared to saline-treated WT group; ∗ p < 0.05 and ∗∗ p < 0.01 compared to saline-treated ob/ob group.

Article Snippet: The primary antibodies against phospho-AMPK (p-AMPK), AMPK, phospho-acetyl-CoA carboxylase (p-ACC), ACC, peroxisome proliferator-activated receptor gamma (PPAR γ ), CCAAT/enhancer-binding protein alpha (C/EBP α ), sterol regulatory element binding protein 1c (SREBP-1c), PPAR γ coactivator 1-alpha (PGC-1 α ), uncoupling protein 2 (UCP2), UCP3, phospho-PI3K (p-PI3K), PI3K, phospho-Akt (p-Akt), Akt, phospho-mammalian target of rapamycin (p-mTOR), mTOR, phospho-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phospho-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), 4EBP1, phospho-forkhead box O3a (p-FoxO3a), FoxO3a, and α -tubulin were purchased from Cell Signaling Technology (Beverly, MA, USA) and used in a 1 : 1000 dilution.

Techniques:

Journal: PLoS ONE

Article Title: Long-Term Artificial Sweetener Acesulfame Potassium Treatment Alters Neurometabolic Functions in C57BL/6J Mice

doi: 10.1371/journal.pone.0070257

Figure Lengend Snippet: Dose-dependent effects of acute ACK exposure upon extracellular acidification rate (ECAR:A, upper panel) and oxygen consumption rate (OCR: A, lower panel) in neuronal cells. 2-DG, (25 mM), was used as a positive control metabolic toxin. (B) Acute ACK and 2-DG effects upon intracellular ATP production (C) were evaluated in SH-SY5Y cells after ACK acute treatment. Acute (15 min) ACK (5–25 mM) or 2-DG (25 mM) exposure-mediated changes in metabolic/survival protein expression in neuronal cells: phospho- (p-Ampk) and non-phosphorylated AMP-activated protein kinase (Ampk) (C); phospho- (p-Acc) and non-phosphorylated Acetyl-CoA carboxylase (Acc) (D); phospho- (p-Akt) and non-phosphorylated v-akt murine thymoma viral oncogene homolog 1 (Akt) (E); phospho- (p-Gsk3β) and non-phosphorylated glycogen synthase kinase 3β (Gsk3β) (F); phospho- (p-Creb) and non-phosphorylated cAMP response element-binding (Creb) (G). (H) ACK-mediated (24 h exposure) effects upon neuronal cell viability. Data from 3 independent experiments are expressed as means ± SEM. One-way ANOVA analysis was performed to evaluate changes in ECAR and OCR of each time point, alterations in intracellular ATP levels and cellular protein expression changes, where the treatment was one independent variable. p<0.05 was considered statistically significant throughout the study. Error bars represent the ±95% confidence interval. *p≤0.05, **p≤0.01, ***p≤0.001.

Article Snippet: Blots were probed with antibodies to phospho-Ser9-Gsk3β, Gsk3β, phospho-Ser473-Akt, Akt, phospho-Ser79-acetyl-coenzyme A carboxylase (p-Acc), acetyl-coenzyme A carboxylase (Acc), phospho-Thr172-Ampk, Ampk, phosphor-Ser133-Creb, Creb, phosphor-Tyr490-TrkB, TrkB, phospho-Erk1/2, Erk1/2, Syp, Syn1,Ppp1r9b and PSD95 (Cell Signaling Technology, Beverly, MA); T1r3 and Bdnf(Santa Cruz Biotechnologies, Dallas, TX).

Techniques: Positive Control, Expressing, Binding Assay

Journal: PLoS ONE

Article Title: Long-Term Artificial Sweetener Acesulfame Potassium Treatment Alters Neurometabolic Functions in C57BL/6J Mice

doi: 10.1371/journal.pone.0070257

Figure Lengend Snippet: (A) ACK-mediated alteration in expression levels of hippocampal proteins associated with energy metabolism were measured using western blot. (B) ACK-induced reduction of hippocampal sweet-taste receptor T1r3subunit (C) ACK-induced reduction of hippocampal glucose transporter 1 (Glut1/Slc2a1). ACK-mediated effects upon expression of Ampk (D), phosphorylated Ampk (p-Ampk: E) were measured to generate a p-Ampk/Ampk ratio (F). ACK-mediated effects upon the expression of Acc (G), phosphorylated Acc (p-Acc: H) were measured to generate a p-Acc/Acc ratio (I). Student’s t-test was used for comparison between control and ACK treatment group. Data are means ± SEM. *p≤0.05, **p≤0.01, ***p≤0.001, n = 3/group.

Article Snippet: Blots were probed with antibodies to phospho-Ser9-Gsk3β, Gsk3β, phospho-Ser473-Akt, Akt, phospho-Ser79-acetyl-coenzyme A carboxylase (p-Acc), acetyl-coenzyme A carboxylase (Acc), phospho-Thr172-Ampk, Ampk, phosphor-Ser133-Creb, Creb, phosphor-Tyr490-TrkB, TrkB, phospho-Erk1/2, Erk1/2, Syp, Syn1,Ppp1r9b and PSD95 (Cell Signaling Technology, Beverly, MA); T1r3 and Bdnf(Santa Cruz Biotechnologies, Dallas, TX).

Techniques: Expressing, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Honokiol activates AMP-activated protein kinase in breast cancer cells via an LKB1-dependent pathway and inhibits breast carcinogenesis

doi: 10.1186/bcr3128

Figure Lengend Snippet: Honokiol activates AMPK and inhibits pS6K and 4EBP1 phosphorylation in breast cancer cells . (a) MCF7 and MDA-MB-231 cells were treated with honokiol (HNK, 2.5 μ M ) for indicated time intervals. U, untreated cells. Total protein was isolated, and equal amounts of proteins were resolved with SDS-PAGE and subjected to immunoblot analysis by using specific antibodies for phosphorylated AMPK (pAMPK-Thr 172) and phosphorylated ACC (pACC). The membranes were reblotted by using total AMPK and ACC antibodies as controls. The blots are representative of multiple independent experiments. The histogram is the mean of densitometric analysis showing relative density units (RDUs) of the Western blot signal for pAMPK and pACC normalized to total AMPK or ACC in three separate experiments. * P < 0.005, compared with untreated controls. (b) Breast cancer cells were treated with honokiol as in (a) and subjected to immunoblot analysis by using specific antibodies for phosphorylated pS6K (p-pS6K) and phosphorylated 4EBP1 (p-4EBP1). The membranes were reblotted by using total pS6K and p-4EBP1 antibodies as controls. The blots are representative of multiple independent experiments. The histogram is the mean of densitometric analysis showing relative density units (RDUs) of the Western blot signal for p-pS6K and p-4EBP1 normalized to total pS6K or 4EBP1 in three separate experiments. * P < 0.001, compared with untreated controls.

Article Snippet: Antibodies for p-AMPK (phospho-AMPK), AMPK, ACC, p-ACC (phospho-ACC), pS6K, p-pS6K (phospho-S6K), 4EBP1, p-4EBP1 (phospho-4EBP1), p-Akt (phospho-Akt), Akt, and LKB1 (3047) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Isolation, SDS Page, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Antiobesity Effect of Astilbe chinensis Franch. et Savet. Extract through Regulation of Adipogenesis and AMP-Activated Protein Kinase Pathways in 3T3-L1 Adipocyte and High-Fat Diet-Induced C57BL/6N Obese Mice

doi: 10.1155/2018/1347612

Figure Lengend Snippet: The effect of Astilbe chinensis Franch. et Savat. (AC) extract on gene expression in 3T3-L1 adipocytes. (a) Expression of AMPK, ACC phosphorylation, (b) PGC-1 α , PPAR α and (c) ATGL, p-HSL mRNA and protein activated in the 3T3-L1 adipocytes and normalized by western blot (upper) and real-time PCR (lower). Data are presented as the mean ± SEM of three independent experiments, each performed in triplicate. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 compared with differentiated cells (DM).

Article Snippet: The primary antibodies PPAR γ , C/EBP α , SREBP1, FAS, SCD-1, phosphor-AMPK (p-AMPK), AMPK, phosphor-ACC (p-ACC), ACC, PGC-1 α , PPAR α , ATGL, phosphor-hormone sensitive lipase (p-HSL), and HSL were purchased form Cell Signaling (Danver, USA).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Antiobesity Effect of Astilbe chinensis Franch. et Savet. Extract through Regulation of Adipogenesis and AMP-Activated Protein Kinase Pathways in 3T3-L1 Adipocyte and High-Fat Diet-Induced C57BL/6N Obese Mice

doi: 10.1155/2018/1347612

Figure Lengend Snippet: Effects of Astilbe chinensis Franch. et Savat. (AC) extract on AMPK and ACC phosphorylation (a), PGC-1 α , PPAR α (b) and ATGL, p-HSL (c) gene expression in the epididymal fat, and determined by western blot (upper) and real-time PCR (lower). ∗∗∗ p < 0.001 compared with HFD mice.

Article Snippet: The primary antibodies PPAR γ , C/EBP α , SREBP1, FAS, SCD-1, phosphor-AMPK (p-AMPK), AMPK, phosphor-ACC (p-ACC), ACC, PGC-1 α , PPAR α , ATGL, phosphor-hormone sensitive lipase (p-HSL), and HSL were purchased form Cell Signaling (Danver, USA).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction