Journal: Metabolism: clinical and experimental

Article Title: Chronic high fat diet induces cardiac hypertrophy and fibrosis in mice

doi: 10.1016/j.metabol.2015.04.010

Figure Lengend Snippet: Effects of HFD and intermittent fasting on the expression of GSK-3β in the left ventricle. A: representative Western blotting. B: quantification of phospho-GSK-3β (P-GSK-3β) at Ser9. C: quantification of phospho-GSK-3β (P-GSK-3β) at Tyr216. D: quantification of GSK-3β protein. Results are means ± S.D. (n = 6 for panels B and C, = 8 for panel D). * P < 0.05 compared with control. RD: regular diet, HFD: high fat diet, IF: intermittent fasting.

Article Snippet: After being blocked with Protein-Free T20 Blocking Buffer (Cat. No. 37573, Thermo Scientific), membranes were incubated with the following primary antibodies: the rabbit monoclonal anti-GSK-3β antibody (1:1000 dilution; catalog number: 9315; Cell Signaling Technology Inc.), the rabbit polyclonal anti-p-GSK-3β (Ser9) antibody (1:500 dilution; catalog number: 9336; Cell Signaling Technology Inc.), the rabbit polyclonal anti-p-GSK-3β (Tyr216) antibody (1:500 dilution; catalog number: sc-135653; Santa Cruz Biotechnology, Santa Cruz, CA), the rabbit monoclonal anti-β-catenin antibody (1:500 dilution; catalog number: 8480; Cell Signaling Technology Inc.), the rabbit polyclonal anti-YAP antibody (1:500 dilution; catalog number: 4912; Cell Signaling Technology Inc., Danvers, MA), the rabbit polyclonal anti-p-YAP (Ser127) antibody (1:500 dilution; catalog number: 4911; Cell Signaling Technology Inc.), the rabbit polyclonal anti-LC3A/B antibody (1:500 dilution; catalog number: 4108; Cell Signaling Technology Inc.), the rabbit polyclonal anti-p62 antibody (1:500 dilution; catalog number: 5114; Cell Signaling Technology Inc.), the rabbit monoclonal anti-cleaved caspase-3 (Asp175) antibody (1:500 dilution; catalog number: 9664; Cell Signaling Technology Inc.), the goat polyclonal anti-GATA4 antibody (1:500 dilution; catalog number: sc-1237; Cell Signaling Technology Inc.), the rabbit polyclonal anti-glyceraldehydes 3-phosphate dehydrogenase (GAPDH) antibody (1:2000 dilution; Catalog number: G9545; Sigma-Aldrich), or the rabbit polyclonal anti-histone H3 antibody (1:3000 dilution; catalog number: 9715; Cell Signaling Technology Inc.).

Techniques: Expressing, Western Blot, Control

Journal: Cell reports

Article Title: Synthetic Essentiality of Metabolic Regulator PDHK1 in PTEN-Deficient Cells and Cancers

doi: 10.1016/j.celrep.2019.07.063

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-p-GSK-3α/β (S21/9) , Cell Signaling Technology , Cat# 9331; RRID: AB_329830.

Techniques: Microarray, Recombinant, DC Protein Assay, Viability Assay, Chromatin Immunoprecipitation, Expressing, Luciferase, Malachite Green Assay, Release Assay, shRNA, Plasmid Preparation, Software

Journal: iScience

Article Title: N6-methyladenosine modification promotes hepatocarcinogenesis through circ-CDYL-enriched and EpCAM-positive liver tumor-initiating exosomes

doi: 10.1016/j.isci.2023.108022

Figure Lengend Snippet:

Article Snippet: rabbit polyclonal anti-p-GSK-3β (Ser9) , Abclonal , Cat#AP0358; RRID: AB_2771152.

Techniques: Control, Virus, Over Expression, Clinical Proteomics, Recombinant, Protease Inhibitor, Western Blot, Purification, Immunoprecipitation, CCK-8 Assay, cDNA Synthesis, Sequencing, Software, Real-time Polymerase Chain Reaction

Journal: International Neurourology Journal

Article Title: PI3K-Akt-Wnt Pathway Is Implicated in Exercise-Induced Improvement of Short-term Memory in Cerebral Palsy Rats

doi: 10.5213/inj.1836224.112

Figure Lengend Snippet: Effects of treadmill exercise on glycogen synthase kinase3β (GSK-3β) and phosphorylated GSK-3β (p-GSK-3β) expressions in the hippocampus. Upper panel: Representative expressions of GSK-3β and p-GSK-3β in the hippocampus. Lower panel: Relative GSK-3β/p-GSK-3β ratio in each group. A, control group; B, exercise group; C, cerebral palsy group; D, cerebral palsy and exercise group. *P<0.05 compared to the control group. # P<0.05 compared to the cerebral palsy group.

Article Snippet: As the primary antibodies, mouse monoclonal anti-Wnt-3 antibody (1:1,000, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), mouse monoclonal anti-PI3K antibody (1:1,000, Santa Cruz Biotechnology), rabbit polyclonal anti-p-PI3K antibody (1:1,000, Santa Cruz Biotechnology), rabbit polyclonal anti-Akt antibody (1:1,000, Cell Signaling Technology Inc., Beverly, Massachusetts, USA), rabbit polyclonal anti-p-Akt antibody (1:1,000, Cell Signaling Technology), rabbit polyclonal anti-GSK-3β antibody (1:1,000, Santa Cruz Biotechnology), rabbit polyclonal anti-p-GSK-3β antibody (1:1,000, Santa Cruz Biotechnology), and mouse monoclonal anti-β-catenin antibody (1:1000, Santa Cruz Biotechnology) were used.

Techniques: Control

Journal: Redox Biology

Article Title: Distinct dual roles of p -Tyr42 RhoA GTPase in tau phosphorylation and ATP citrate lyase activation upon different Aβ concentrations

doi: 10.1016/j.redox.2020.101446

Figure Lengend Snippet: Low concentrations of Aβ induce p -Ser422 tau through RhoA/ROCK/GSK3β signalling. ( A and B ) Several types of p-Tau were detected with Western blot using specific antibodies after stimulating HT22 cells with soluble (s) or fibrillary (f) form of Aβ for short (Aβ25-35) or long (Aβ1-42) peptides (1 μM, 24 h). In one case, NGF (100 ng/ml) was treated for 72 h, and several types of p-Tau were assessed (one-way ANOVA, p*<0.05, **<0.01, ***<0.001). ( C ) Tat-C3 (Rho inhibitor, 1 μg/ml) and Y27532 (ROCK inhibitor, 10 μM) were pretreated for 1 h, and then fAβ (1 μM) was treated for 24 h to HT22 cells. P-Ser422 Tau, p -Tyr216 GSK-3β and RhoA were detected by Western blot (one-way ANOVA, p*<0.05, **<0.01, ***<0.001, ****<0.0001). (D) GTP-RhoA levels of HT22 cells were measured using a pulldown assay with GST-RBD after fAβ (1 μM) treatment for the given times, (E) or after various concentration of fAβ (1 μM) treatment for 24 h. (F – G) Immunofluorescence of a vehicle- and sAβ (10 μM or 100 μM)-injected CA1 and CA2 regions of C57BL/6 mice brains (4 months of age) was revealed with anti-NeuN (neuronal nucleus), p -Ser422 Tau, AT8 ( p -Ser202/Thr205 Tau) and -p-Y42 Rho antibodies.

Article Snippet: Anti-p-S389 GSK3β (14850-1-AP) was obtained Proteintech (Manchester, UK).

Techniques: Western Blot, Concentration Assay, Immunofluorescence, Injection

Journal: Redox Biology

Article Title: Distinct dual roles of p -Tyr42 RhoA GTPase in tau phosphorylation and ATP citrate lyase activation upon different Aβ concentrations

doi: 10.1016/j.redox.2020.101446

Figure Lengend Snippet: AD patient specimens also show similar features of the Aβ signalling pathway in HT22 cells. (A) Immunohistochemistry for p-Y42 Rho in the CA1, CA2, CA3, CA4, DG, and EC regions of the hippocampus of control normal persons and AD patients (Scale bar = 50 μm). (B) RNA expression of RhoA in temporal cortex of normal and AD brain was analysed. ( C,D ) Several proteins likely related to Aβ signalling pathway were analysed with western blotting in brain tissues of normal controls and AD patients (C) and relative intensity of protein bands was quantified. (unaired, two-tailed t -test, p* < 0.05). (D) . (E,F) Expression of mRNA levels in normal and AD patients were analysed by transcriptome analysis: NADPH oxidase components (E) and enzymes for fatty acid and lipid synthesis. Data are means of 6–9 independent samples ± SE (unaired, two-tailed t -test, p** <0.01) (E,F) . (G) Schematic illustration showing two Aβ-dependent concentration signalling pathways: low concentrations of Aβ (1 μM) can activate RhoA/ROCK/p-Tyr216 GSK-3β, causing p -Ser422 tau and neuronal damage. On the other hand, when p -Tyr216 GSK-3β (active form) is phosphorylated and p -SREBP1 is ubiquitinated and degraded, a transcription factor to induce enzymes for lipid synthesis including ACL, a decrease of ACL expression is seen. High concentrations of Aβ (10 μM) produces enhanced levels of ROS and phosphorylates p38 MAPK (T180/Y182), which can inactivate GSK-3β by phosphorylating Ser389 and, in turn, activates SREBP1, leading to an increase of ACL level as well as fatty acid synthesis and cellular senescence. High ROS cause DNA damage which activates ATM by phosphorylating Ser1981 then phosphorylates ACL Ser455. High concentration of Aβ (10 μM) also facilitates nuclear translocation of p -Tyr42 Rho, associating with the promoter region of NADK gene, thereby likely resulting in high level of NADP and NADPH. Finally, NADPH can be a hydrogen donor for FS activity as a defence mechanism rather than NADPH oxidase activity that induces highly ROS, thereby leading to cell death.

Article Snippet: Anti-p-S389 GSK3β (14850-1-AP) was obtained Proteintech (Manchester, UK).

Techniques: Immunohistochemistry, Control, RNA Expression, Western Blot, Two Tailed Test, Expressing, Concentration Assay, Translocation Assay, Activity Assay