difluorophenacetyl l alanyl s phenylglycine 2 butyl ester code named dapt hy 13027 mce  (MedChemExpress)

Journal: Physiological Reports

Article Title: Glutamine enhances endothelial cell survival and vasodilation by increasing glutathione to reduce oxidative stress

doi: 10.14814/phy2.70737

Figure Lengend Snippet: Glutamine increased vasodilation ex vivo but did not impact eNOS phosphorylation or nitric oxide in vitro. (a) Mouse carotid artery vasodilation in response to acetylcholine with an intact endothelium. Isolated mouse carotid arteries were incubated at 4°C overnight in physiological saline containing either 0 or 2 mM glutamine. Vasodilation was measured the following day by pressure myography. n = 4–5 arteries per condition. (b) representative Western blot with quantification of eNOS and p‐eNOS in HCAEC cultured in 0 and 2 mM glutamine. n = 10–11 samples per condition. (c) HCAECs were treated with 0, 0.5, and 2 mM glutamine in 5.5 and 15 mM glucose. L‐glucose at 2 mM and 11.5 mM was included as positive control for glutamine and glucose respectively. After 24 h cells were treated with 20 μM of yoda1 and then the nitrites in the culture media was measured using a Griess assay. n = 9 samples. Data shown as mean ± standard deviation. Statistical significance determined by repeated measures ANOVA (a), Mann–Whitney nonparametric test (b), and two‐way ANOVA followed by Tukey's post hoc test.

Article Snippet: Endothelial‐independent vasodilation was measured by preincubating carotid arteries with eNOS inhibitor NG‐Nitroarginine methyl ester hydrochloride (L‐NAME, 10 −5 M; HY‐18729A; Med Chem Express) for 1 h. Vasodilation to acetylcholine was then measured as previously described.

Techniques: Ex Vivo, Phospho-proteomics, In Vitro, Isolation, Incubation, Saline, Western Blot, Cell Culture, Positive Control, Griess Assay, Standard Deviation, MANN-WHITNEY

Journal: Communications Biology

Article Title: GDF11 alleviates glucocorticoid-induced osteonecrosis of the femoral head by regulating angiogenesis via the PI3K-AKT-eNOS pathway

doi: 10.1038/s42003-025-09078-5

Figure Lengend Snippet: A Microarray heatmap of HUVECs from the MP groups with or without GDF11 treatment (n = 3 per group). B Bubble plots of differentiated pathways in indicated groups. C GO enrichment analysis of DEGs was conducted comparing MP groups with versus without GDF11 supplementation. D GSEA demonstrated significant positive enrichment in gene sets involved in cell migration, circulatory system process, and blood vessel morphogenesis after GDF11 treatment. E , F Immunofluorescence staining of p-PI3K and p-eNOS in HUVECs, with quantitative analysis of mean fluorescence intensity in control (untreated), MP-treated, and MP + GDF11-treated groups (n = 3 per group). Scale bar = 20 μm. G Western blotting was performed to detect PI3K, AKT, eNOS, p-PI3K, p-AKT, and p-eNOS protein expression in control (untreated), MP-treated, and MP + GDF11-treated groups. H Quantification of Western blotting analysis in different groups (n = 3 per group). I NO levels in control (untreated), MP-treated, and MP + GDF11-treated groups (n = 3 per group). Significant differences are indicated as follows: ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: HUVECs were treated with MP (100 μM) or MP (100 μM) + GDF11 (10 ng/ml) in the presence or absence of the PI3K inhibitor LY294002 (10 μM, A10547, Adooq, China) or eNOS inhibitor L-NAME (100 μM, HY-18729A, MCE, USA).

Techniques: Microarray, Migration, Immunofluorescence, Staining, Fluorescence, Control, Western Blot, Expressing

Journal: Communications Biology

Article Title: GDF11 alleviates glucocorticoid-induced osteonecrosis of the femoral head by regulating angiogenesis via the PI3K-AKT-eNOS pathway

doi: 10.1038/s42003-025-09078-5

Figure Lengend Snippet: A Western blotting was performed to detect expression of PI3K, AKT, eNOS, p-PI3K, p-AKT, and p-eNOS in the following groups: MP, MP + GDF11-treated, MP + GDF11 + LY294002-treated, and MP + GDF11 + L-NAME-treated cells. B Quantification of Western blotting analysis in different groups (n = 3 per group). C NO levels in MP, MP + GDF11-treated, MP + GDF11 + LY294002-treated, and MP + GDF11 + L-NAME-treated groups (n = 3 per group). D Representative immunofluorescence images of MP, MP + GDF11-treated, MP + GDF11 + LY294002-treated, and MP + GDF11 + L-NAME-treated groups showing red (CD31), green (phalloidine), blue (nuclei). Scale bar = 20 μm. E Quantitative analysis of mean fluorescence in different groups (n = 3 per group). F Western blotting was performed to detect expression of CD31 and VEGFA in the following groups: MP, MP + GDF11-treated, MP + GDF11 + LY294002-treated, and MP + GDF11 + L-NAME-treated cells. G Quantification of Western blotting analysis in different groups (n = 3 per group). H , I Scratch assay for 0, 12, 24, and 48 h and quantitative analysis in different groups (n = 3 per group). Scale bar = 200 μm. J Transwell assays were performed in four groups: MP, MP + GDF11-treated, MP + GDF11 + LY294002-treated, and MP + GDF11 + L-NAME-treated cells. Scale bar = 100 μm. K Quantification of migration cells of Transwell assay (n = 3 per group). L Tube formation assays were performed in four groups: MP, MP + GDF11-treated, MP + GDF11 + LY294002-treated, and MP + GDF11 + L-NAME-treated cells. Scale bar = 200 μm. M – O Quantification of Tube formation assay (n = 3 per group). Significant differences are indicated as follows: ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: HUVECs were treated with MP (100 μM) or MP (100 μM) + GDF11 (10 ng/ml) in the presence or absence of the PI3K inhibitor LY294002 (10 μM, A10547, Adooq, China) or eNOS inhibitor L-NAME (100 μM, HY-18729A, MCE, USA).

Techniques: Western Blot, Expressing, Immunofluorescence, Fluorescence, Wound Healing Assay, Migration, Transwell Assay, Tube Formation Assay