human enos nos3 small interfering rna  (Santa Cruz Biotechnology)

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: NOSTRIN is an emerging negative regulator of decompensated cirrhotic patients with portal hypertension.

doi: 10.1016/j.dld.2024.08.050

Figure Lengend Snippet: Fig. 3. (A, i) Pathological changes observed by H&E of human cirrhotic liver and control liver sections (magnification - 10X). The cirrhotic patient’s liver showed increased Nostrin expression and decreased peNOS expression, which was concomitant with reduced NO bioavailability. Representative images (magnification - 20X) of human cirrhotic liver tissue sections showing elevated hepatic Nostrin levels by DAB immunohistochemistry (A, ii). Representative images (magnification - 20X) of human liver tissue sections showing elevated hepatic eNOS levels in the control liver compared to the cirrhotic liver by DAB immunohistochemistry A, iii). Hepatic Nostrin protein expression by immunoblot assay with densitometry quantification (B) and mRNA expression by qPCR (C). Liver total eNOS and p-eNOS expression were evaluated by immunoblot assay with densitometry quantification (D). Hepatic total and phosphorylated eNOS/eNOS in the serine 1177 residue (peNOS) expression by immunoblot assay with densitometry quantification (D) and mRNA expression by qPCR (E). The hepatic cGMP levels were measured by ELISA (F). Data are expressed as mean ± standard error of mean (SEM). Symbols represent ∗p < 0.05; ∗∗p < 0.01 compared to control liver. β-actin was used as an internal loading control.

Article Snippet: Tissues were incubated with 5 % goat s t s ( t o r a H m ( t c s 2 f a a u R t e R A i ( A a w r i P t m 2 t p ( s t ( p R a t ( t i w i a ( H t a 2 t s a s S t l ( P 3 3 a c w t i t s p o B 3 a a erum for 1 hr and incubated overnight at 4 °C with primary anibodies against rabbit polyclonal anti-human Nostrin (1:100; cat: c-134,803, Santacruz) and rabbit polyclonal anti- human eNOS 1:100; cat: PA-1712–1, Bosterbio).

Techniques: Control, Expressing, Immunohistochemistry, Western Blot, Residue, Enzyme-linked Immunosorbent Assay

Journal: Current Research in Physiology

Article Title: The comparison of endothelial function of moderate intensity interval exercise with continuous exercise in healthy men

doi: 10.1016/j.crphys.2022.07.003

Figure Lengend Snippet: Comparison of vascular markers between intermittent and continuous exercise.

Article Snippet: Then, the enzyme-linked immunosorbent assay (ELISA) method was used for determination of serum concentrations of N-terminal proANP (NTproANP), N-terminal proBNP (NTproBNP), N-terminal proCNP (NTproCNP), endothelial nitric oxide synthase activity, endothelin 1 (ET-1), adiponectin, and leptin (Elabscience Cat. No: E-EL-H1848, Cat. No: E-EL-H0902, Cat. No: E-EL-H2538, Cat. No: E-EL-H0755, Cat. No: E-EL-H0064, Cat. No: E-EL-H0004, Cat. No: E-EL-H0113 respectively).

Techniques: Comparison, Activity Assay

Journal: The Journal of investigative dermatology

Article Title: Loss of Endothelial Cell Matrix Metalloproteinase 14 Reduces Melanoma Growth and Metastasis by Increasing Tumor Vessel Stability.

doi: 10.1016/j.jid.2021.12.016

Figure Lengend Snippet: Figure 6. eNOS expression and NO production in EC. (a) Analysis of DAF-FM fluorescence intensity of primary ECs measured at 495 nm. Scatter dot plot shows the mean SD; n ¼ 3. **P < 0.005. (b) NOS3 transcript amplification in primary ECs of Mmp14fl/fland Mmp14EC‒/‒ mice using S26 as control. Scatter dot plot shows the mean SD; n ¼ 9; *P < 0.05. (c) Primary ECs stained for eNOS (red) and nuclei (DAPI, blue). eNOS expression intensity was quantified. The scatter dot plot shows the mean SD. **P < 0.005. Bar ¼ 50 mm. (d) Quantitative analysis of eNOS and MMP14 expression in human melanoma. Intensities are arbitrarily set as þ (low), þþ (moderate), and þþþ (high). Graphs show the percentage of eNOS expression intensity in low, moderate, or high MMP14- expressing vessels. Arrowheads indicate representative vessels. n ¼ 21. Bar ¼ 250 mm. DAF-FM, diaminofluorescein-FM; EC, endothelial cell; eNOS, endothelial nitric oxide synthase; MMP14, matrix metalloproteinase 14; NO, nitric oxide.

Article Snippet: Human umbilical vein endothelial cells were transfected with 10 nM human MMP14 small interfering RNA (Ambion, 8877, Thermo Fisher Scientific) and human eNOS (NOS3) small interfering RNA (sc-36093, Santa Cruz Biotechnology).

Techniques: Expressing, Control, Staining

Journal: EXCLI Journal

Article Title: Crocetin promotes angiogenesis in human endothelial cells through PI3K-Akt-eNOS signaling pathway

doi: 10.17179/excli2019-1175

Figure Lengend Snippet: The molecular formula of Crocetin (A). Relative viability measured by WST-1 in HUVECs exposed to various concentrations of Crocetin over a period of 72 h (B) (n=12). Measuring HUVECs viability in the combined regime of Crocetin, Wortmannin (a PI3K inhibitor) and NMA (an eNOS inhibitor) (C). Data are expressed as mean ± SD (n=12). *p<0.05, **p<0.01, ***p<0.001 using one-way ANOVA with Tukey post-hoc test.

Article Snippet: Briefly, we transferred 100 μl rabbit anti-human VEGFR-1 (Dilution: 1 μg/ml; Cat no: ab32152; Abcam), rabbit anti-human VEGFR-2 (Dilution: 1 μg/ml; Cat no: ab39256; Abcam), mouse anti-human eNOS (Cat no: sc-376542, Santa Cruz Biotechnology) and mouse anti-human phosphor eNOS (Cat no: sc-81510, Santa Cruz Biotechnology) into each well of polystyrene treated 96-well plate (SPL) and kept them overnight at 4 °C.

Techniques:

Journal: EXCLI Journal

Article Title: Crocetin promotes angiogenesis in human endothelial cells through PI3K-Akt-eNOS signaling pathway

doi: 10.17179/excli2019-1175

Figure Lengend Snippet: Gelatin zymogram showing MMP-9 and MMP-2 activities in HUVECs treated with Crocetin (A) (n=3). RT-PCR analysis of VEGF expression (B) (n=3). Western blot analysis of p-Akt/Akt ratio (C). Western blotting revealed the non-significant increase of p-Akt/Akt ratio during the 45 min-incubation of HUVECs with 50 µM Crocetin and these effects were diminished from time points 60 to 120 min. (n=3). Measuring the p-Akt/Akt and p-eNOS/eNOS ratios by ELISA (D, E, and F) (n=12) by using One-way ANOVA and Tukey's Post Hoc. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: Briefly, we transferred 100 μl rabbit anti-human VEGFR-1 (Dilution: 1 μg/ml; Cat no: ab32152; Abcam), rabbit anti-human VEGFR-2 (Dilution: 1 μg/ml; Cat no: ab39256; Abcam), mouse anti-human eNOS (Cat no: sc-376542, Santa Cruz Biotechnology) and mouse anti-human phosphor eNOS (Cat no: sc-81510, Santa Cruz Biotechnology) into each well of polystyrene treated 96-well plate (SPL) and kept them overnight at 4 °C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Incubation, Enzyme-linked Immunosorbent Assay

Journal: EXCLI Journal

Article Title: Crocetin promotes angiogenesis in human endothelial cells through PI3K-Akt-eNOS signaling pathway

doi: 10.17179/excli2019-1175

Figure Lengend Snippet: The molecular formula of Crocetin (A). Relative viability measured by WST-1 in HUVECs exposed to various concentrations of Crocetin over a period of 72 h (B) (n=12). Measuring HUVECs viability in the combined regime of Crocetin, Wortmannin (a PI3K inhibitor) and NMA (an eNOS inhibitor) (C). Data are expressed as mean ± SD (n=12). *p<0.05, **p<0.01, ***p<0.001 using one-way ANOVA with Tukey post-hoc test.

Article Snippet: Briefly, we transferred 100 μl rabbit anti-human VEGFR-1 (Dilution: 1 μg/ml; Cat no: ab32152; Abcam), rabbit anti-human VEGFR-2 (Dilution: 1 μg/ml; Cat no: ab39256; Abcam), mouse anti-human eNOS (Cat no: sc-376542, Santa Cruz Biotechnology) and mouse anti-human phosphor eNOS (Cat no: sc-81510, Santa Cruz Biotechnology) into each well of polystyrene treated 96-well plate (SPL) and kept them overnight at 4 °C.

Techniques:

Journal: EXCLI Journal

Article Title: Crocetin promotes angiogenesis in human endothelial cells through PI3K-Akt-eNOS signaling pathway

doi: 10.17179/excli2019-1175

Figure Lengend Snippet: Gelatin zymogram showing MMP-9 and MMP-2 activities in HUVECs treated with Crocetin (A) (n=3). RT-PCR analysis of VEGF expression (B) (n=3). Western blot analysis of p-Akt/Akt ratio (C). Western blotting revealed the non-significant increase of p-Akt/Akt ratio during the 45 min-incubation of HUVECs with 50 µM Crocetin and these effects were diminished from time points 60 to 120 min. (n=3). Measuring the p-Akt/Akt and p-eNOS/eNOS ratios by ELISA (D, E, and F) (n=12) by using One-way ANOVA and Tukey's Post Hoc. * p <0.05; ** p <0.01; *** p <0.001.

Article Snippet: Briefly, we transferred 100 μl rabbit anti-human VEGFR-1 (Dilution: 1 μg/ml; Cat no: ab32152; Abcam), rabbit anti-human VEGFR-2 (Dilution: 1 μg/ml; Cat no: ab39256; Abcam), mouse anti-human eNOS (Cat no: sc-376542, Santa Cruz Biotechnology) and mouse anti-human phosphor eNOS (Cat no: sc-81510, Santa Cruz Biotechnology) into each well of polystyrene treated 96-well plate (SPL) and kept them overnight at 4 °C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Incubation, Enzyme-linked Immunosorbent Assay