Journal: bioRxiv

Article Title: Post-ischemic inactivation of HIF prolyl hydroxylases in endothelium promotes maladaptive kidney repair by inducing glycolysis

doi: 10.1101/2023.10.03.560700

Figure Lengend Snippet: ( A ) Experimental scheme for HPAECs subjected to 0.5% O 2 for 18 hours in the presence of syrosingopine (5 µM) or MCT4 siRNA followed by reoxygenation for 8 hours in the presence of IL-1β (1 ng/ml). ( B ) mRNA levels of VCAM1 and ICAM1 in syrosingopine- vs vehicle-treated HPAECs, that were activated by Hypoxia/Reoxygenation and IL-1β. ( C ) THP1 monocyte adhesion to inflamed ECs. THP1 monocyte cells, labeled with Green CMFDA Dye, were introduced on a monolayer of HPAECs that had been subjected to the indicated experimental conditions. Following a 90-minute incubation period, floating cells were washed away and adhered THP1 cells were visualized using a fluorescent microscope and subsequently quantified. Representative images of fluorescent THP1 cells attached to ECs in different experimental groups are presented. Scale bar, 200 μm. ( D ) mRNA expression of VCAM1 and ICAM1 in HPAECs transfected with control or MCT4 siRNA and subjected to Hypoxia/Reoxygenation and IL-1β ( E ) THP1 monocyte adhesion to inflamed ECs. Labeled THP1 cells were introduced on a monolayer of HPAECs that had been subjected to the indicated experimental conditions. Following incubation, adhered THP1 cells were visualized quantified as in C . Representative images of fluorescent THP1 cells attached to ECs in different experimental conditions are presented. Scale bar, 200 μm. Data are represented as mean ± SEM. Statistics were determined by one-way ANOVA with Sidak correction for multiple comparisons. n=3-4; *, P < 0.05; **, P <0.01; ***, P <0.001; ****, P < 0.0001; ns, not statistically significant. Nx, Normoxia; Hx, Hypoxia/Reoxygenation; Veh, vehicle; Syro, syrosingopine; C, negative control siRNA; MCT4si, MCT4siRNA.

Article Snippet: Human primary pulmonary artery ECs (HPAECs) were obtained from ATCC and grown on gelatin-coated dishes in Endothelial Cell Basal Medium-2 (Lonza, Catalog # CC-3156) supplemented with EGM-2 SingleQuots Supplements (Lonza, Catalog # CC-4176).

Techniques: Labeling, Incubation, Microscopy, Expressing, Transfection, Control, Negative Control

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Repurposing rosiglitazone, a PPAR-γ agonist and oral antidiabetic, as an inhaled formulation, for the treatment of PAH

doi: 10.1016/j.jconrel.2018.04.049

Figure Lengend Snippet: The levels of eNOS expression in human PAH PAECs in hypoxic and normoxic condition (A), PPARγ and endothelin-1 in PAH-ECs (B); PPARγ and NOX-4 expression in PAH-SMCs (C). n=3.

Article Snippet: 2.2.10 The effects of rosiglitazone on human PAH-ECs and PAH-SMCs We grew human pulmonary arterials ECs and SMCs (collected from PAH patients), upon receipt from the Pulmonary Hypertension Breakthrough Initiative (PHBI), in gelatin coated dishes using MV2 (PromoCell GmbH, Heidelberg, Germany) and SmGM (Lonza Inc., Williamsport, PA) media, respectively for 5-7 days until their confluency.

Techniques: Expressing

Journal:

Article Title: Design and Synthesis of Biomimetic Hydrogel Scaffolds with Controlled Organization of Cyclic RGD Peptides

doi: 10.1021/bc800441v

Figure Lengend Snippet: 2D seeding and culturing of endothelial cells (ECs) on hydrogels. (a, b) Phase contrast of ECs 2 h and 24 h after seeding on PEGDA hydrogel, respectively; (c, d) Phase contrast of ECs 2 h and 24 h after seeding on cRGD-PEGDA hydrogel, respectively. The images show that ECs seeded on cRGD-PEGDA hydrogels exhibited higher initial cell attachment, greater cell spreading, and higher cell density than on PEGDA hydrogels. (Scale bar = 100 µm).

Article Snippet: Cell culturing on hydrogels Human pulmonary artery endothelial cells (ECs) (passage 7, Cambrex) were grown to 80–90% confluence using complete growth medium (CGM) in 25-cm 2 tissue culture flasks (Costar) at 37°C and 5% CO 2 .

Techniques: Cell Attachment Assay