Journal: Journal of Animal Science and Biotechnology
Article Title: Maternal supply of cysteamine alleviates oxidative stress and enhances angiogenesis in porcine placenta
Figure Lengend Snippet: Cysteamine (CS) pretreatment prolongs the phosphorylation of Stat3 in H 2 O 2 -treated PVECs. A , B Western blotting analysis of the expression of Phospho-Stat3 (p-Stat3), NADPH oxidase 2 (NOX2), and vascular endothelial growth factor A (VEGF-A). Cells were pretreated with various concentrations of cysteamine (0.5, 1 or 2 mmol/L CS) for 2 h, and then challenged with H 2 O 2 (200 μmol/L) for 24 h ( n = 3). C , D Representative images of tube formation by PVECs, pretreated with CS (0.5 mmol/L) and/or inhibitors of Stat3 (5 μmol/L stattic) for 2 h, and then challenged with H 2 O 2 (200 μmol/L) for 24 h ( n = 5; bar = 100 μm)5. E CCK8 assay was used to measure cell viability after different treatments as described above ( n = 6). F , G Western blotting analysis of the expression of p-Stat3 and VEGF-A ( n = 3). Data are presented as mean ± SEM. Different letters indicate significant differences at P < 0.05
Article Snippet: After blocking with TBS/T buffer containing 5% milk, the membranes were incubated with the primary antibodies against vascular endothelial growth factor A (VEGF-A) (19003–1-AP, Proteintech, USA, 1:1,000), NADPH oxidase 2 (NOX2) (19013–1-AP, Proteintech, USA, 1:1,000), signal transducer and activator of transcription-3 (Stat3)(ab76315, Abcam, USA, 1:1,500), p-Stat3 (ab68153, Abcam, USA, 1:1,500), and β-actin (4970, CST, USA, 1:1,000).
Techniques: Western Blot, Expressing, CCK-8 Assay