akt phosphorylation inhibitor ly294002  (MedChemExpress)

Journal: World Journal of Diabetes

Article Title: Fractional carbon dioxide laser-induced photothermal activation of mesenchymal stem cell-derived exosomes accelerates diabetic wound healing by enhancing angiogenesis

doi: 10.4239/wjd.v17.i1.112942

Figure Lengend Snippet: LY294002 suppresses exosomes derived from CO 2 laser-preconditioned adipose-derived mesenchymal stem cells -induced angiogenesis. b P < 0.01; c P < 0.001. Data are presented as mean ± SD ( n = 3). A and B: Western blot analysis of phosphorylated protein kinase B, protein kinase B, hypoxia-inducible factor-1α, and vascular endothelial growth factor-A protein expression levels in human umbilical vein endothelial cells (HUVECs) treated with LY294002 vs negative control and LY294002 + exosomes derived from CO 2 laser-preconditioned adipose-derived mesenchymal stem cells vs exosomes derived from CO 2 laser-preconditioned adipose-derived mesenchymal stem cells; C and D: 5-Ethynyl-2’-deoxyuridine incorporation assay to assess HUVEC proliferation. Scale bar = 100 μm; E: Cell Counting Kit-8 assay to evaluate HUVEC viability; F and G: Transwell migration assay to assess the migratory capacity of HUVECs. Scale bar = 200 μm; H and I: In vitro wound healing assay to evaluate cell migration. Scale bar = 200 μm; J and K: Tube formation assay to examine the formation of capillary-like structures by HUVECs. Scale bar = 200 μm. EdU: 5-Ethynyl-2’-deoxyuridine; NC: Negative control; CO 2 laser-Exos: Exosomes derived from CO 2 laser-preconditioned adipose-derived mesenchymal stem cells; AKT: Protein kinase B; p-AKT: Phosphorylated protein kinase B; HIF-1α: Hypoxia-inducible factor-1α; VEGF-A: Vascular endothelial growth factor-A.

Article Snippet: To evaluate the involvement of the PI3K/AKT signaling pathway in CO 2 laser-Exos-mediated angiogenesis, HUVECs were pretreated with either the AKT phosphorylation inhibitor LY294002 or the activator SC79 (MedChem Express, United States).

Techniques: Derivative Assay, Western Blot, Expressing, Negative Control, Cell Counting, Transwell Migration Assay, In Vitro, Wound Healing Assay, Migration, Tube Formation Assay

Journal: Burns & Trauma

Article Title: Sphingosine-1-phosphate derived from PRP-Exos promotes angiogenesis in diabetic wound healing via the S1PR1/AKT/FN1 signalling pathway

doi: 10.1093/burnst/tkad003

Figure Lengend Snippet: PRP-Exos-S1P regulates FN1 via the AKT signalling pathway. ( a – d ) The relative levels of FN1, p-AKT and VEGF-A in HUVECs after different treatments were measured by western blotting in the LY294002 vs NC and LY294002 + PRP-Exos vs PRP-Exos groups. ( e – h ) The relative levels of FN1, p-AKT and VEGF-A in HUVECs after different treatments were measured by western blotting in the SC79 vs NC and SC79 + shS1PR1 vs shS1PR1 groups. ( i , j ) Immunofluorescence for FN1 was measured in the LY294002 vs NC and LY294002 + PRP-Exos vs PRP-Exos groups. ( k , l ) Immunofluorescence for FN1 was measured in the SC79 vs NC and SC79 + shS1PR1 vs shS1PR1 groups. * * * p < 0.001, * * p < 0.01, * p < 0.05. LY294002 inhibitor of AKT phosphorylation, SC79 agonist of AKT phosphorylation, NC normal control, PRP platelet-rich plasm, PRPExos exosomes derived from PRP, FN1 fibronectin 1, p-AKT phosphorylated protein kinase B, t-AKT total protein kinase B, VEGF-A vascular endothelial growth factor A

Article Snippet: The AKT phosphorylation inhibitor LY294002 (S1005, Selleck) and agonist SC79 (S7863, Selleck) were used to study the PI3K/AKT signalling pathway.

Techniques: Western Blot, Immunofluorescence, Phospho-proteomics, Control, Derivative Assay

Journal: PLoS ONE

Article Title: Long-Time Treatment by Low-Dose N -Acetyl-L-Cysteine Enhances Proinflammatory Cytokine Expressions in LPS-Stimulated Macrophages

doi: 10.1371/journal.pone.0087229

Figure Lengend Snippet: RAW264.7 cells were treated with 2/ml LPS was added into the conditioned media, and the cells were incubated for further 10 minutes (A, B). The cell lysates were subjected to western blot analysis using antibodies against pAKT, AKT, pERK1/2, ERK1/2 (A), pp38, p38, pJNK1/2, JNKs, or GAPDH (B). Ratios of AKT, ERK1/2 (A), p38 and JNK (B) phosphorylation relative to protein expression levels were photographically calculated using Image-J software (Image processing program). The relative phosphorylation levels in comparison to 0 h NAC treatment (set as 1.0) are shown in numbers. C. The pAP1-luc plasmid was stably transfected into RAW264.7 cells. The established cells were exposed to 2 mM NAC for 0 hour, 1 hour, 20 hours or 30 hours, and then the cells were stimulated with 100 ng/ml LPS for further 4 hours. Luciferase activities in the cell lysates were normalized by the protein contents. * p <0.05 versus control.

Article Snippet: U0126, a specific inhibitor of ERK activation pathway, SB203580, a specific p38 kinase inhibitor, SP 600125, a specific JNK inhibitor and LY294002, and a specific inhibitor of AKT phosphorylation were from CALBIOCHEM (San Diego, CA).

Techniques: Incubation, Western Blot, Expressing, Software, Plasmid Preparation, Stable Transfection, Transfection, Luciferase

Journal: BioMed Research International

Article Title: High Glucose Promotes CD36 Expression by Upregulating Peroxisome Proliferator-Activated Receptor γ Levels to Exacerbate Lipid Deposition in Renal Tubular Cells

doi: 10.1155/2017/1414070

Figure Lengend Snippet: PPAR γ is upregulated by the HG-promoted AKT phosphorylation, which can regulate CD36 expression in HK-2 cells. HK-2 cells were cultured with NG (5.6 mM), NG + LY294002 (15 uM), HG (30 mM), or HG + LY294002 (15 uM) for 48 h. P-AKT, AKT, and PPAR γ were examined by western blotting (a, b). HK-2 cells were pretreated with an agonist (RSG, 5 uM)/antagonist (GW9662, 2.5 uM) of PPAR γ for 1 h, followed by 48 h of NG (5.6 uM)/HG (30 uM) stimulation for the analysis of CD36 protein levels. Using western blotting, cell lysates were analyzed (c, d). All experiments were repeated thrice. Band intensities were normalized to β -actin band intensity using densitometry. The data were represented as the means ± SD. ∗ P < 0.05 versus NG; # P < 0.05 versus HG.

Article Snippet: LY294002, an inhibitor of AKT phosphorylation, was purchased from Beyotime Institute of Biotechnology (Haimeng, China).

Techniques: Expressing, Cell Culture, Western Blot

Journal: BioMed Research International

Article Title: High Glucose Promotes CD36 Expression by Upregulating Peroxisome Proliferator-Activated Receptor γ Levels to Exacerbate Lipid Deposition in Renal Tubular Cells

doi: 10.1155/2017/1414070

Figure Lengend Snippet: The roles of AKT-PPAR γ signaling pathway and CD36 in regulating HG-induced lipid accumulation in HK-2 cells. The HK-2 cells were transfected with CD36siRNA or nontargeting control siRNA (NTC siRNA) under HG (30 uM) and NG (5.6 uM) conditions and western blot analysis was used to determine CD36siRNA-mediated knockdown of CD36 in the HK-2 cells. Band intensities were normalized to β -actin band intensity using densitometry. The data from three independent experiments were represented as means ± SD (a). ∗ P < 0.05 versus NTCsiRNA. By using Oil Red O staining, lipid content was detected in the HK-2 cells incubated with NG (5.6 mM), NG + NTCsiRNA, NG + CD36siRNA, NG + LY294002 (15 uM), NG + RSG (5 uM), NG + GW9662 (2.5 uM), HG (30 mM), HG + NTCsiRNA, HG + CD36siRNA, HG + LY294002 (15 uM), HG + RSG (5 uM), and HG + GW9662 (2.5 uM) for 48 h, respectively (b). Relative levels of lipid content were quantified by measuring relative absorbance of the eluted Oil Red O dye at 500 nm. Data from three independent experiments were represented as means ± SD (c). ∗ P < 0.05 versus NG; # P < 0.05 versus HG.

Article Snippet: LY294002, an inhibitor of AKT phosphorylation, was purchased from Beyotime Institute of Biotechnology (Haimeng, China).

Techniques: Transfection, Western Blot, Staining, Incubation

Journal: BioMed Research International

Article Title: Shenmai Injection Supresses Glycolysis and Enhances Cisplatin Cytotoxicity in Cisplatin-Resistant A549/DDP Cells via the AKT-mTOR-c-Myc Signaling Pathway

doi: 10.1155/2020/9243681

Figure Lengend Snippet: SMI enhances cisplatin cytotoxicity in cisplatin-resistant A549/DDP cells via the AKT-mTOR-c-Myc signaling pathway. (a) LY294002 (PI3K inhibitor), (b) rapamycin (mTOR inhibitor), and (c) 10058-F4 (c-Myc inhibitor) were treated or not treated in cisplatin-resistant A549/DDP cells via western blot analysis of protein expression levels of genes involved in cell glucose metabolism.

Article Snippet: LY294002 (phosphorylated Akt inhibitor; Abmole Bioscience, Shanghai, China; 20 μ M), rapamycin (phosphorylated mTOR inhibitor; Abmole Bioscience, Shanghai, China; 5 nM), and 10058-F4 (c-Myc inhibitor; Abmole Bioscience, Shanghai, China; 20 μ M) were added to the cells for 12 h, and the same volume of dimethyl sulfoxide (DMSO) was used as the control.

Techniques: Western Blot, Expressing