Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Biatractylolide Modulates PI3K-Akt-GSK3 β -Dependent Pathways to Protect against Glutamate-Induced Cell Damage in PC12 and SH-SY5Y Cells

doi: 10.1155/2017/1291458

Figure Lengend Snippet: Effects of biatractylolide on Akt and Gsk3β expressions of PC12 and SH-SY5Y cells induced by glutamate . (a)-(b) represent protein characterization of Akt and Gsk3 β after PC12 and SH-SY5Y cells were treated with various concentrations of biatractylolide for 30 min before glutamate treatment for 24 hours. Blots were also probed for β -actin as loading controls. (c)-(d) showed the ratio of different proteins to β -actin was calculated by the band density of each cell line using Image J software. ∗ p < 0.05 and ∗∗ p < 0.01 versus control group; # p < 0.05, ## p < 0.01, and ### p < 0.005 versus model group.

Article Snippet: Antibodies for the protein characteristics were against total Akt, phosphor-Akt, total GSK3 β , phosphor-GSK3 β , and β -actin which were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Software

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Interleukin-4 Boosts Insulin-Induced Energy Deposits by Enhancing Glucose Uptake and Lipogenesis in Hepatocytes

doi: 10.1155/2018/6923187

Figure Lengend Snippet: IL-4 performs synergistic activity to boost insulin signaling, promoting insulin-triggered hepatic glucose uptake, glycogen synthesis, and fatty acid synthesis. HepG2 and Huh7 cells were serum starved, followed by IL-4, insulin (INS), or combined treatment (IL-4 + INS) for the indicated time. Cell lysates were harvested, with p-AKT, p-GSK-3 α / β , and GLUT2 analyzed by Western blotting (a). Hepatic glucose uptake and glycogen synthesis (b). PEPCK (c) p-ACC1, FAS, and DGAT2 (d) analyzed by Western blotting. Bar graphs showed the corresponding quantitative results ( n = 3). ∗ p < 0.05 vs. 0 min; ∗∗ p < 0.01 vs. 0 min; # p < 0.05 vs. INS at the same time point.

Article Snippet: Reagents were obtained from the following sources: antibodies against Akt, phospho-Ser473 Akt, GSK-3 α / β , and phosphor-GSK-3 α / β Ser21/9 from Cell Signaling Technology (Danvers, MA, USA); anti-ACC1 and anti-DGAT2 from Gentex Inc. (Irvine, CA, USA); anti-IL-4R from Abcam (San Francisco, CA, USA); mouse IL-4 from Millipore (Temecula, CA, USA); ECL reagent from Calbiochem (Merck Millipore, Billerica, MA, USA); insulin, anti-GATA3, anti- β -actin, fatty acid uptake, and glycogen assay kits from Sigma (St. Louis, MO, USA); anti-phospho-STAT-6 from Millipore Corporation; anti-SREBP-1, anti-PPAR α , anti-GAPDH, anti-GLUT2, and anti-PEPCK from Santa Cruz Biotechnology Inc.; TRIzol Reagent from Life Technologies (Carlsbad, CA, USA); anti-FAS from BD Biosciences; triglyceride quantification kit from BioVision Inc. (Milpitas, CA, USA).

Techniques: Activity Assay, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Interleukin-4 Boosts Insulin-Induced Energy Deposits by Enhancing Glucose Uptake and Lipogenesis in Hepatocytes

doi: 10.1155/2018/6923187

Figure Lengend Snippet: Regulation of hepatic glucose and lipid metabolism by IL-4. IL-4 stimulates AKT and GSK-3 α / β phosphorylations, which in turn upregulates glucose uptake and GS activity for promoting hepatic glycogen synthesis. In addition, IL-4 promotes hepatic triglyceride contents through enhancing FFA uptake and the expression/activity of lipogenic enzymes, including ACC, FAS, and DGAT2. The net effects of IL-4 on hepatocytes are to promote energy storage by enhancing insulin-stimulated glucose uptake and lipid synthesis under physiological condition.

Article Snippet: Reagents were obtained from the following sources: antibodies against Akt, phospho-Ser473 Akt, GSK-3 α / β , and phosphor-GSK-3 α / β Ser21/9 from Cell Signaling Technology (Danvers, MA, USA); anti-ACC1 and anti-DGAT2 from Gentex Inc. (Irvine, CA, USA); anti-IL-4R from Abcam (San Francisco, CA, USA); mouse IL-4 from Millipore (Temecula, CA, USA); ECL reagent from Calbiochem (Merck Millipore, Billerica, MA, USA); insulin, anti-GATA3, anti- β -actin, fatty acid uptake, and glycogen assay kits from Sigma (St. Louis, MO, USA); anti-phospho-STAT-6 from Millipore Corporation; anti-SREBP-1, anti-PPAR α , anti-GAPDH, anti-GLUT2, and anti-PEPCK from Santa Cruz Biotechnology Inc.; TRIzol Reagent from Life Technologies (Carlsbad, CA, USA); anti-FAS from BD Biosciences; triglyceride quantification kit from BioVision Inc. (Milpitas, CA, USA).

Techniques: Activity Assay, Expressing

Journal: Frontiers in Immunology

Article Title: A Requirement of Protein Geranylgeranylation for Chemokine Receptor Signaling and Th17 Cell Function in an Animal Model of Multiple Sclerosis

doi: 10.3389/fimmu.2021.641188

Figure Lengend Snippet: Protein geranylgeranylation is required for heterotrimeric small G-protein mediated GPCR-proximal signaling. (A) Western blot analysis of CCL20-induced phosphorylation of Akt and Erk in Th17 cells and density of phosphor-Akt calculated using ImageJ; (B) FTase I and GGTase I scores of the 12 γ-subunits of the small heterotrimeric GTPases calculated using an algorithm described in the text; (C) Fold change of the expression of the genes encoding the 12 γ-subunits of heterotrimeric small GTPase in naive and effector CD4 + T cells analyzed by qRT-PCR; (D) Flow cytometry analysis of EGFP expression in Th17 cells infected with retrovirus carrying cDNAs encoding mutant γ-subunits capable of being farnesylated; (E) Western blot analysis of phosphor-Akt and phosphor-Gsk3βS9 in Th17 cells described in (D) ; (F) Density of phosphor-Akt and phosphor-Gsk3β in (E) calculated using Image J (Results are representatives of three biologically independent experiments, n.s. statistically not significant; ** p < 0.01, *** p < 0. 001, **** p < 0. 0001unpaired t -test).

Article Snippet: Protein samples were resolved on SDS-PAGE gels and subjected by immunoblot with the following antibodies: Pggt1b (Sigma HPA030646), npRap1a (Santa Cruz sc1482), Phospho-Akt S473 (Cell Signaling, 4060), phosphor-Gsk3β Ser9 (Cell Signaling 9322), phosphor-Erk (Cell Signaling, 9101), phosphor-Stat3 Tyr705 (Cell Signaling, 9145), phosphor-Smad2 (Cell Signaling 3108), phosphor-Smad3 (Cell Signaling, 9520), and β-Actin-HRP (Sigma, A3854).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Flow Cytometry, Infection, Mutagenesis