ser 79 phosphorylated acc  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: AMP-Activated Protein Kinase (AMPK) Mediates Nutrient Regulation of Thioredoxin-Interacting Protein (TXNIP) in Pancreatic Beta-Cells

doi: 10.1371/journal.pone.0028804

Figure Lengend Snippet: (A) Effects of non-metabolizable MEDICA analogs on AMPK activity and TXNIP expression. MEDICA analogs were generated by substitution of alpha, omega-dicarboxylic acids of C16 chain length. INS-1E cells were incubated for 16 h at 11.1 (G11.1) and 22.2 (G22.2) mmol/l glucose with and without beta, beta'- or alpha, alpha'-tetramethyl-hexadecanedioic acid (M16alpha/alpha and M16beta/beta, respectively). AMPK activity was assessed by measuring phosphorylated acetyl CoA carboxylase (pACC). pACC and TXNIP protein levels were analyzed by Western blot. Quantification of TXNIP expression normalized to GAPDH (B) and ACC phosphorylation normalized to ACC (C) presented. (n = 3). (D) Effect of palmitate treatment on AMPK activity. INS-1E cells were incubated at different glucose concentrations without and with 0.5 mmol/l palmitate in KRBH buffer for 1 h. A representative gel of phospho- and total ACC and GAPDH is shown, and quantification of ACC phosphorylation presented. Results are expressed as fold of untreated cells at 3.3 mmol/l glucose (n = 10). * p<0.05, ** p<0.01, ∧ p<0.001 for the difference between MEDICA analog-treated cells and controls at the same glucose concentration (B, C), between untreated cells at different glucose concentrations and controls at 3.3 mmol/l glucose (G3.3) (D), and between palmitate-treated cells and the paired controls at the same glucose concentration (D).

Article Snippet: Protein expression in whole cell and nuclear extracts was studied by Western blot using antibodies against TXNIP (MBL International Co, Woburn, MA), total AMPK, total and phosphorylated (Ser 79) acetyl CoA carboxylase (ACC) (Cell Signaling Technology, Beverley, MA), AMPKalpha1 and alpha2 (Upstate Biotechnology, Lake Placid, NY), ChREBP, Lamin B and GAPDH (Santa Cruz Biotechnology).

Techniques: Activity Assay, Expressing, Generated, Incubation, Western Blot, Concentration Assay

Journal: Phytomedicine Plus

Article Title: Berberine is as effective as the anti-obesity drug Orlistat in ameliorating betel-nut induced dyslipidemia and oxidative stress in mice

doi: 10.1016/j.phyplu.2021.100098

Figure Lengend Snippet: Fig. 6. Berberine and Orlistat restored phosphorylation of ACC at Ser-79 in AEBN-treated mice; a- histograms showing ratio of protein intensity of pACC (Ser-79) to total ACC in the liver of male and female mice; b- representative western blots immuno-probed for pACC (Ser-79) and total ACC with blots immuno-probed for β-actin serving as loading controls in the liver of male and female mice respectively. Data are expressed as mean ± SD of three indepen dent experiments. The p-values have been determined using one way ANOVA followed by Tukey’s Multiple Comparison Test; n = 30 mice per treatment group. Western blots are representative of 3 independent experiments.

Article Snippet: Polyvinyl Difluoride (PVDF) membrane was obtained from (Sigma Aldrich, St. Louis USA); antibodies against total and phosphorylated AMPK (Thr-172) (# 2603S and #2535S respectively) total and phosphorylated ACC (Ser-79) (# 3676S and #11818S respectively), FASN (#3180) and Beta- actin (# 4970S) were obtained from Cell Signaling Technology (Massachusetts, United States).

Techniques: Phospho-proteomics, Western Blot, Comparison

Journal: Molecular Metabolism

Article Title: Deletion of intestinal epithelial AMP-activated protein kinase alters distal colon permeability but not glucose homeostasis

doi: 10.1016/j.molmet.2021.101183

Figure Lengend Snippet: Inducible deletion of gut AMPK did not exacerbate the development of HFD-induced obesity and metabolic dysfunctions . (A) Experimental timeline for tamoxifen treatment and control diet (CD) or high-fat diet (HFD) challenge for 16 weeks in male WT and i-IEC AMPK KO mice. (B) Western blotting analysis of AMPKα, phospho-AMPKα-Thr172, ACC, and phospho-ACC-Ser79 expression in intestinal epithelial cells isolated from the ileum of WT and i-IEC AMPK KO mice at the completion of CD or HFD challenge. β-actin was used as a loading control. (C) Body weight monitoring in WT and i-IEC AMPK KO mice on CD or HFD for 16 weeks. n = 7–13 mice. (D) Body weight and body composition examination (expressed as a percentage of fat mass and lean mass relative to total body mass) in WT and i-IEC AMPK KO mice at the end of CD or HFD challenge. n = 7–13 mice. (E) Blood glucose profile during glucose tolerance tests in WT and i-IEC AMPK KO mice on CD or HFD. AUC, area under the curve was calculated. n = 6–11 mice. All of the data are expressed as means ± SEM. Statistical analysis was performed by two-way repeated measures ANOVA or two-way ANOVA with Bonferroni's post hoc test. ##p < 0.01 and ###p < 0.001 indicate a diet effect in WT mice. §§p < 0.01 and §§§p < 0.001 indicate a diet effect in i-IEC AMPK KO mice. $p < 0.05 and $$$p < 0.001 indicate a diet effect within genotype. CD, control diet; HFD, high-fat diet; black bars, WT mice; dashed bars, i-IEC AMPK KO mice.

Article Snippet: Primary antibodies directed against total AMPKα (#2532), AMPKβ1 (#4178), AMPKβ2 (#4148), AMPKγ2 (#2536), AMPKα phosphorylated at Thr-172 (#2531), total acetyl-CoA carboxylase 1/2 (ACC1/2) (#3676), ACC phosphorylated at Ser-79 (#3661), and β-actin (#4967) were purchased from Cell Signaling Technology.

Techniques: Control, Western Blot, Expressing, Isolation

Journal: Scientific Reports

Article Title: Induction of glucose uptake in skeletal muscle by central leptin is mediated by muscle β 2 -adrenergic receptor but not by AMPK

doi: 10.1038/s41598-017-15548-6

Figure Lengend Snippet: Effects of leptin on insulin signalling and AMPK activity in soleus muscle of WT and β-less mice. Representative immunoblot analysis of phosphorylated (p) and total forms of IR ( a ), Akt ( b ), and the α subunit of AMPK ( c ) in soleus or Gastro-W at 6 h after injection of saline (−) or leptin (+) into the VMH of WT or β-less mice is shown together with quantitation of the corresponding pIR/IR, pAkt/Akt, and pAMPKα/AMPKα ratios. Representative data were shown in duplicate. Representative immunoblots for α-tubulin were also shown in. ( b ) Quantitative data are expressed as a percentage of the corresponding value for injection of saline into WT mice and are means ± S.E.M. ( n = 4). * P < 0.05 versus corresponding value for saline injection into WT mice; ‡ P < 0.05 versus corresponding value for saline injection into β-less mice (one-way ANOVA and Bonferroni’s multiple-range test).

Article Snippet: Primary antibodies included those to the Tyr 1146 -phosphorylated β subunit of IR, to Ser 473 -phosphorylated Akt, to the Thr 172 -phosphorylated form of the α subunit of AMPK, to Ser 79 -phosphorylated acetyl-CoA carboxylase (ACC), and to total forms of these various proteins and α-tubulin (Cell Signalling Technology).

Techniques: Activity Assay, Western Blot, Injection, Saline, Quantitation Assay