Journal: Journal of Inflammation (London, England)

Article Title: Differential expression of nitric oxide synthases in porcine aortic endothelial cells during LPS-induced apoptosis

doi: 10.1186/1476-9255-9-47

Figure Lengend Snippet: Forward and reverse primer sequences, RT-PCR product length and accession number (Acc.No.) in the EMBL database

Article Snippet: The membranes were then incubated overnight at 4°C with a 1:200 dilution of an anti-iNOS (610332, BD Transduction) rabbit polyclonal antibody in PBS-T20 with 3% milk powder or at 4°C overnight with a 1:500 dilution of an anti-eNOS mouse monoclonal antibody (Sa-258, Biomol, Butler Pike, Plymouth Meeting, PA, USA) in Tris Buffered Saline-T20 (TBS-T20 20 mM Tris–HCl, pH 7.4, 500 mM NaCl, 0.1% T-20).

Techniques: Sequencing

Journal: Journal of Inflammation (London, England)

Article Title: Differential expression of nitric oxide synthases in porcine aortic endothelial cells during LPS-induced apoptosis

doi: 10.1186/1476-9255-9-47

Figure Lengend Snippet: Relative gene expression of iNOS and eNOS in pAECs under standard culture conditions at different time periods. Relative mRNA data are expressed as delta Ct (HPRT Ct - iNOS or eNOS Ct). The data represent the mean ± SEM (n = 3). Time of culture did not influence iNOS and eNOS mRNA expression.

Article Snippet: The membranes were then incubated overnight at 4°C with a 1:200 dilution of an anti-iNOS (610332, BD Transduction) rabbit polyclonal antibody in PBS-T20 with 3% milk powder or at 4°C overnight with a 1:500 dilution of an anti-eNOS mouse monoclonal antibody (Sa-258, Biomol, Butler Pike, Plymouth Meeting, PA, USA) in Tris Buffered Saline-T20 (TBS-T20 20 mM Tris–HCl, pH 7.4, 500 mM NaCl, 0.1% T-20).

Techniques: Gene Expression, Expressing

Journal: Journal of Inflammation (London, England)

Article Title: Differential expression of nitric oxide synthases in porcine aortic endothelial cells during LPS-induced apoptosis

doi: 10.1186/1476-9255-9-47

Figure Lengend Snippet: Relative gene expression of iNOS (A) and eNOS (B) in pAECs treated with LPS (10 μg/ml) at different time periods (n = 3). LPS increased iNOS mRNA expression while eNOS mRNA expression was decreased. Relative mRNA data are expressed as the fold of increase (ΔΔCt method) in respect to the control (CTR = mean ± SEM of all control time points). Error bars represent the range of relative expression. The different letters above the bars indicate significant differences in the various time points (p < 0.05, ANOVA post hoc Tukey’s test).

Article Snippet: The membranes were then incubated overnight at 4°C with a 1:200 dilution of an anti-iNOS (610332, BD Transduction) rabbit polyclonal antibody in PBS-T20 with 3% milk powder or at 4°C overnight with a 1:500 dilution of an anti-eNOS mouse monoclonal antibody (Sa-258, Biomol, Butler Pike, Plymouth Meeting, PA, USA) in Tris Buffered Saline-T20 (TBS-T20 20 mM Tris–HCl, pH 7.4, 500 mM NaCl, 0.1% T-20).

Techniques: Gene Expression, Expressing, Control

Journal: Journal of Inflammation (London, England)

Article Title: Differential expression of nitric oxide synthases in porcine aortic endothelial cells during LPS-induced apoptosis

doi: 10.1186/1476-9255-9-47

Figure Lengend Snippet: Expression of eNOS protein in pAECs treated with LPS (10 μg/ml) for different time periods (7, 15, 24 h). A ) LPS induced a significant decrease of eNOS protein expression in respect to the control (CTR = mean ± SEM of all control time points). The data represent the mean ± SEM (n = 3) of the relative protein content (AU = Arbitrary Units). The different letters above the bars indicate significant differences among the various time points (p < 0.05, ANOVA post hoc Tukey’s test, n = 3). B ) Representative Western Blot of eNOS and relative housekeeping β-tubulin were reported.

Article Snippet: The membranes were then incubated overnight at 4°C with a 1:200 dilution of an anti-iNOS (610332, BD Transduction) rabbit polyclonal antibody in PBS-T20 with 3% milk powder or at 4°C overnight with a 1:500 dilution of an anti-eNOS mouse monoclonal antibody (Sa-258, Biomol, Butler Pike, Plymouth Meeting, PA, USA) in Tris Buffered Saline-T20 (TBS-T20 20 mM Tris–HCl, pH 7.4, 500 mM NaCl, 0.1% T-20).

Techniques: Expressing, Control, Western Blot

Journal: Journal of Inflammation (London, England)

Article Title: Differential expression of nitric oxide synthases in porcine aortic endothelial cells during LPS-induced apoptosis

doi: 10.1186/1476-9255-9-47

Figure Lengend Snippet: Representative eNOS immunofluorescent staining on pAEC. Green fluorescence indicates positive immunostaining revealed by FITC, red fluorescence indicates nuclear staining with Propidium Iodide (magnification 400X). A , B ) eNOS protein localisation in standard cultured pAECs; a cytoplasmatic signal evenly diffused was detected. ( C - F ) LPS treatment resulted in a modification of the signal with more intense staining in the perinuclear region.

Article Snippet: The membranes were then incubated overnight at 4°C with a 1:200 dilution of an anti-iNOS (610332, BD Transduction) rabbit polyclonal antibody in PBS-T20 with 3% milk powder or at 4°C overnight with a 1:500 dilution of an anti-eNOS mouse monoclonal antibody (Sa-258, Biomol, Butler Pike, Plymouth Meeting, PA, USA) in Tris Buffered Saline-T20 (TBS-T20 20 mM Tris–HCl, pH 7.4, 500 mM NaCl, 0.1% T-20).

Techniques: Staining, Fluorescence, Immunostaining, Cell Culture, Modification

Journal: Scientific reports

Article Title: Chronic stress in adulthood results in microvascular dysfunction and subsequent depressive-like behavior.

doi: 10.1038/s41598-024-74902-7

Figure Lengend Snippet: Fig. 4. Microvascular dysfunction induced by endothelial injury can cause depression-like behavior. (A) Timeline of the experiment in eNOS+/ − mice and control littermates. We selected three time periods of 2 months, 4 months, and 6 months as experimental nodes. (B) Standardized statistical results of cerebral PU in eNOS+/− mice and WT. (C) Standardized statistical results of right forepaw PU in eNOS+/− mice and WT. (D) Standardized statistical results of right hindpaw PU in eNOS+/− mice and WT. (E) Standardized statistical results of PORH time in eNOS+/− mice and WT. (F) Standardized statistical results of sucrose preference during SPT. (G) Standardized statistical results of immobility time during FST. (H) Trend plots of cerebral perfusion and depression-like behavioral. eNOS: endothelial nitric oxide synthase, WT: wild type littermates. WT group was used as the standard value (100%) in each group. P*<0.05, P**<0.01, P***<0.001 vs. WT.

Article Snippet: Male endothelial nitric oxide synthase knockout (eNOS−/−) mice were obtained from The Jackson Laboratory (stock NO.002684).

Techniques: Control

Journal: Scientific reports

Article Title: Chronic stress in adulthood results in microvascular dysfunction and subsequent depressive-like behavior.

doi: 10.1038/s41598-024-74902-7

Figure Lengend Snippet: Fig. 5. Arginase inhibitors rescued microvascular dysfunction and depression-like behavior caused by endothelial injury in eNOS+/−mice. (A) Timeline of drug treatment experiments in eNOS+/− mice and WT littermates. Laser Doppler Measurement and behavioral test were performed before and 6 weeks after intraperitoneal injection of arginase inhibitor and saline from the age of 4 months. (B) Right hindpaw PU of eNOS+/− mice and WT mice during treatment. (C) Sucrose preference during treatment. (D) Percentage of immobility time to total time during FST after treatment. (E) Cerebral PU after treatment. (F) Time of PORH after treatment. (G) Nitric oxide in brain tissue after treatment. Nor-NOHA: N-Hydroxy-nor-L-arginine Dihydrochloride, Typical arginase inhibitors. P*<0.05, P**<0.01, P***<0.001 vs. WT. N = 8.

Article Snippet: Male endothelial nitric oxide synthase knockout (eNOS−/−) mice were obtained from The Jackson Laboratory (stock NO.002684).

Techniques: Injection, Saline