antiphosphorylated phospho enos ser1177  (Cell Signaling Technology Inc)

Journal: Journal of pharmacological sciences

Article Title: Comparison of cytotoxicity of cigarette smoke extract derived from heat-not-burn and combustion cigarettes in human vascular endothelial cells.

doi: 10.1016/j.jphs.2021.07.005

Figure Lengend Snippet: Fig. 4. Effects of crude and n/t-free hCSE/bCSE on eNOS phosphorylation at Ser1177 and total eNOS protein expression in human endothelial EA.hy926 cells. Crude and n/t-free cigarette smoke of 42 puffs of Ploom S, glo, IQOS, and hi-lite was bubbled into 5 mL ice-cold KrebseHEPES solution to prepare individual 100% CSE solution (P42 hCSE for Ploom S, G42 hCSE for glo, I42 hCSE for IQOS, and H42 bCSE for hi-lite). Cells were treated with crude or n/t-free hCSE/bCSE at indicated concentrations for 4 h. (A, D, G, and J) Panels are representative immunoblots with anti-phosphorylated (phospho)-eNOS (Ser1177), anti-total (phospho and nonphosphorylated)-eNOS (t-eNOS), and anti-GAPDH antibodies used to determine levels of phospho-eNOS, t-eNOS, and GAPDH, respectively. (B, E, H, and K) Ordinates indicate changes in relative ratio of expression levels of phospho-eNOS to total GAPDH (phospho-eNOS/GAPDH) in the absence or presence of CSE. The p-eNOS/GAPDH ratio was normalized to ratio of control cells (considered as 100%). Data are mean ± SEM of results of four independent experiments (**P < 0.01, compared with control cells; #P < 0.05 and ##P < 0.01, compared with corresponding crude CSE). When no error bar is shown, the error is smaller than the symbol. (C, F, I, and L) Ordinates indicate changes in relative ratio of t-eNOS levels to the total GAPDH expression levels (t-eNOS/GAPDH) in the absence or presence of CSE. The t-eNOS/GAPDH ratio was normalized to ratio of control cells (100%). Data are means ± SEM of results of four independent experiments (*P < 0.05 and **P < 0.01, compared with control cells). When no error bar is shown, the error is smaller than the symbol.

Article Snippet: Antiphosphorylated (phospho)-eNOS (Ser1177) and anti-total eNOS antibodies were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control

Journal: Cardiology Research and Practice

Article Title: Elevated GTP Cyclohydrolase I Pathway in Endothelial Progenitor Cells of Overweight Premenopausal Women

doi: 10.1155/2020/5914916

Figure Lengend Snippet: The GTPCH I/BH4 pathway and the phosphorylation of eNOS in circulating EPCs in the four groups. The level of GTPCH I (a) or intracellular BH4 (b) of EPCs in normal-weight and overweight premenopausal women was higher than that in normal-weight and overweight men. There was no difference in the level of GTPCH I (a) or intracellular BH4 (b) of EPCs between normal-weight and overweight premenopausal women. No significant difference was found in either eNOS phosphorylation (c) or eNOS protein expression (d) of EPCs between the four groups. (e) Representative photographs of GTPCH I, phosphorylated eNOS and eNOS expression of EPCs. Data are given as mean ± SD. ∗ P < 0.05 vs. the same gender group of normal weight; # P < 0.05 vs. the same weight class of premenopausal women.

Article Snippet: Rabbit antiphosphorylated eNOS and anti-eNOS (1 : 1000; Cell Signaling Technology Inc.) and anti-GTPCH I and β -actin (1 : 1000; Santa Cruz Biotechnology Inc.) were used to evaluate eNOS and GTPCH I expression as previously described [ , , ].

Techniques: Phospho-proteomics, Expressing