Journal: Scientific Reports

Article Title: Activated ROCK/Akt/eNOS and ET-1/ERK pathways in 5-fluorouracil-induced cardiotoxicity: modulation by simvastatin

doi: 10.1038/s41598-020-71531-8

Figure Lengend Snippet: Effect of 5-FU and Sim treatments on cardiac tissue contents of ( A ) p -Akt and ( B ) p -eNOS. Values are presented as the mean of 6–7 experiments ± SEM. Statistical analysis was carried out using one-way ANOVA followed by Tukey’s post-hoc test ; as compared to normal (*), Sim (#), and 5-FU (@)-treated groups (p < 0.05). 5-FU 5-fluorouracil, p-Akt (Ser473) phosphorylated protein kinase B at Ser 473, p-eNOS (Ser1177) phosphorylated endothelial nitric oxide synthase at Ser 1177, p-ERK1/2 (Thr202/Tyr204) phosphorylated extracellular signal-regulated kinase 1/2 at Thr 202 & Tyr 204, Sim simvastatin.

Article Snippet: The corresponding ELISA kits were used to determine the cardiac contents of p -NF-κB p65 (Ser536) (Abcam, Cambridge, UK; Cat# ab176647), p -ERK1/2 (Thr202/Tyr204) (Invitrogen, CA, USA; Cat # EMS2ERKP), p -Akt (Ser473) (Elabscience, Wuhan, China; Cat # E-EL-R1135) and p -eNOS (Ser1177) (Cell Signaling Technology, MA, USA; Cat# 7980C).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Chronic Insulin Infusion Down-Regulates Circulating and Urinary Nitric Oxide (NO) Levels Despite Molecular Changes in the Kidney Predicting Greater Endothelial NO Synthase Activity in Mice

doi: 10.3390/ijms19102880

Figure Lengend Snippet: Western blotting for endothelial nitric oxide synthase (eNOS) protein—( A ) Representative Western blots of cortex homogenates. Separate blots were probed for p-S1177-eNOS, p-Y657-eNOS, and total eNOS. Equal amounts of protein were loaded in each lane. Summaries of band densities (normalized to β-actin) for ( B ) p-S1177-eNOS, p-Y657-eNOS, total eNOS; ( C ) Representative western blots of outer medulla homogenates. Summaries of band densities (normalized to β-actin) for ( D ) p-S1177-eNOS, p-Y657-eNOS, total eNOS; ∗ indicates a significant difference between groups by one-way ANOVA (mean ± sem; n = 8–12/group for statistics).

Article Snippet: Antibodies used included: β-actin (monoclonal mouse, Sigma-Aldrich, St. Louis, MO, USA), eNOS (NOS3 (monoclonal rabbit D9A5L, Cell Signaling, Danvers, MA, USA) phosphorylated (serine 1177)-eNOS (monoclonal rabbit C9C3, Cell Signaling), and phosphorylated (Tyrosine 657)-eNOS (NP4031, ECM Biosciences, Versailles, KY, USA).

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Chronic Insulin Infusion Down-Regulates Circulating and Urinary Nitric Oxide (NO) Levels Despite Molecular Changes in the Kidney Predicting Greater Endothelial NO Synthase Activity in Mice

doi: 10.3390/ijms19102880

Figure Lengend Snippet: Western blot of eNOS from unreduced samples—( A ) Representative Western blot for eNOS run on cortex homogenates solubilized with Laemmli without dithiotrietol (DTT) and run under non-reducing conditions. Arrows indicate coupled (active, 280 kDa) and uncoupled (140 kDa) bands. ( B ) Band densities summary; * indicates a significant difference between groups by one-way ANOVA ( p < 0.05, mean ± sem; n = 8/group for statistics).

Article Snippet: Antibodies used included: β-actin (monoclonal mouse, Sigma-Aldrich, St. Louis, MO, USA), eNOS (NOS3 (monoclonal rabbit D9A5L, Cell Signaling, Danvers, MA, USA) phosphorylated (serine 1177)-eNOS (monoclonal rabbit C9C3, Cell Signaling), and phosphorylated (Tyrosine 657)-eNOS (NP4031, ECM Biosciences, Versailles, KY, USA).

Techniques: Western Blot

Journal: Neurochemical Research

Article Title: Prostaglandin E2 EP4 Receptor Activation Attenuates Neuroinflammation and Early Brain Injury Induced by Subarachnoid Hemorrhage in Rats

doi: 10.1007/s11064-016-2168-6

Figure Lengend Snippet: Effects of EP4 receptor on the expression of p-eNOS. Western blot analysis showed AE1-329 increased the expression of Ser1177 p-eNOS 24 h after SAH and AE3-208 showed opposite effects ( a, b ). Data were expressed as mean ± SEM (n = 8, ### p < 0.001 vs. Sham, *p < 0.05 vs. SAH treated with vehicle, **p < 0.01 vs. SAH treated with vehicle)

Article Snippet: Samples were supplemented with 2% beta-mercaptoethanol and 50 mM DTT and boiled for about 5 min. Proteins were separated on tris-glycine 4–15% acrylamide gels and transferred to PVDF membranes soaked in 5% nonfat milk in PBS-Tween 20 (0.05%) for 2 h. The proteins of total caspase-3, cleaved caspase-3, Ser1177 phosphorylated endothelial nitric oxide synthase (Ser1177 p-eNOS) and β-actin were tested by rabbit monoclonal antibody against caspase-3 (Santa Cruz, 1:500), rabbit polyclonal antibody against cleaved caspase-3 (Biorbyt, 1:500), rabbit polyclonal antibody against Ser1177 p-eNOS (Invitrogen, 1:400) and rabbit polyclonal antibody against β-actin (Santa Cruz, 1:500) respectively.

Techniques: Expressing, Western Blot