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