Figures index : Protective Effect of Wheat Derived Non-specific lipid-transfer Protein 2 on Vascular Endothelium Inflammation : Science and Education Publishing

Figures index

Journal of Food and Nutrition Research. 2018, 6(6), 386-392 doi:10.12691/jfnr-6-6-6

Figure 1. Effect of nsLTP2 on cell adhesion molecule expression in HUVECs treated with TNF-α. After 10 nM nsLTP2 (LTP2) pre-treatment for 24 h, HUVECs were stressed with 20 ng/mL TNF-α. Cell lysates were subjected to SDS-PAGE and Western blotting analysis, using primary antibodies directed to VCAM-1, ICAM-1. Images show a representative experiment of three independent analyses. Densitometric analysis of bands intensity, normalized to actin, was expressed as % of stressed cells and was carried out with the ChemiDoc MP System (BIORAD). Data represent means ± SD of three independent experiments. ^§p < 0.05, vs control HUVECs; *p < 0.05, vs HUVECs stressed with TNF-α

Figure 2. Effect of nsLTP2 on HO-1 protein expression level in HUVECs. After treatment with 10 nM nsLTP2 for 24 h, cells lysates were subjected to SDS-PAGE and Western blotting analysis of HO-1 protein expression levels. The image shows a representative experiment of three independent analyses. Densitometric analysis of HO-1 expression, normalized to actin, is expressed as fold increase over control and was carried out with the ChemiDoc MP System (BIORAD). Data represent means ± SD of three independent experiments. §p < 0.05, vs control HUVECs

Figure 3. Immunofluorescence staining of HO-1 in HUVECs after 10 nM nsLTP2 treatment for 24 h After treatment with 10 nM nsLTP2, HUVECs were immunolabeled with anti-HO-1 antibody and relative fluorescent FITCH-conjugated secondary antibody then visualized using immunofluorescence microscopy (Zeiss Axio Scope.A1, magnification 100x). Nuclei were stained with DAPI

Figure 4. Effect of nsLTP2 (LTP2), Atorvastatin and Simvastatin on HO-1 protein expression level. HUVECs were treated with 10 nM nsLTP2 (LTP2), 10 nM - 50 µM Atorvastatin (A) or 10 nM - 50 µM Simvastatin (S) for 24 h. Cell lysates were subjected to SDS-PAGE and Western blotting analysis of HO-1 protein expression level. Densitometric analysis of HO-1 expression, normalized to actin, is expressed as fold increase over control and was carried out with the ChemiDoc MP System (BIORAD). Data represent means ± SD of three independent experiments. §p < 0.05, compared with control HUVECs

Figure 5. Effect of nsLTP2 on VCAM-1 expression in HUVECs treated with TNF-α in the presence or absence of HO-1 inhibitors. HUVECs were treated with 10 nM nsLTP in the presence or absence of HO-1 inhibitors (10nM ZnPP IX and SnPP IX) and subsequently treated with 20ng/mL TNF-α. Cell lysates were subjected to SDS-PAGE and Western blotting analysis of VCAM protein expression level. Densitometric analysis of VCAM expression normalized to actin is expressed as % of stressed cells and was carried out with the ChemiDoc MP System (BIORAD). Data represent means ± SD of three independent experiments. ^§p < 0.05, compared with control HUVECs, *p < 0.05, compared with HUVECs stressed by TNF-α.