primary human paecs  (PromoCell)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Hypoxia Combined With Interleukin‐17 Regulates Hypoxia‐Inducible Factor‐1α/Endothelial Nitric Oxide Synthase Expression in Pulmonary Artery Endothelial Cells

doi: 10.1111/jcmm.70289

Figure Lengend Snippet: HIF‐1α and eNOS levels in pulmonary artery endothelial cells under different experimental conditions. (A, B) Western blot analysis of HIF‐1α and eNOS levels in PAECs. (C–E) The ratio of greyscale values of target bands and β‐Actin bands. (F) Real‐time polymerase chain reaction of the mRNA expression of HIF‐1α in PAECs. Data are expressed as the mean ± standard deviation. * p < 0.05, ** p < 0.01; ns: No statistical differences.

Article Snippet: Human PAECs (ScienCell Research Laboratories, Carlsbad, CA, USA) were cultured in endothelial cell medium (ECM) (ScienCell) with the addition of 5% foetal bovine serum at 37°C with 5% CO 2 .

Techniques: Western Blot, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

Journal: Cells

Article Title: Hypoxia-Induced Mitochondrial ROS and Function in Pulmonary Arterial Endothelial Cells

doi: 10.3390/cells13211807

Figure Lengend Snippet: Hypoxia increases the formation of ROS in human pulmonary artery endothelial cells, and mitochondrial complex I and complex III are responsible for ROS production. ( A ) Exposure to hypoxia for 10 min significantly increased the formation of H 2 O 2 in human PAECs. Cells were incubated with Amplex UltraRed (50 µM) for 20 min. The fluorescence produced by Amplex UltraRed was measured using the FlexStation III reader as an indicator of H 2 O 2 production. Data are from three different experiments and are expressed as mean ± S.E.M. * p < 0.05 compared to normoxia, n = 4. ( B ) Cells were transfected with HyPer for 2 days, treated for 10 min without (control) and with rotenone (10 µM) or myxothiazol (10 μM), and then exposed to hypoxia. The bar graph illustrates that hypoxia enhanced the formation of H 2 O 2 in human PAECs and rotenone and myxothiazol blocked this response, suggesting a role of complex I and complex III in this phenomenon. HyPer-derived fluorescence was measured using an LSM510 confocal microscope. Data are expressed as the media ± S.E.M. and were obtained from at least 50 cells in each group. * p < 0.05 compared to normoxia, n = 5.

Article Snippet: Primary human PAECs were purchased from Lonza (Allendale, NJ, USA) and cultured in Lonza’s EGM-2 media with growth factor (CC-3162 and CC-3129) in a humidified atmosphere of 5% CO 2 and 95% air at 37 °C.

Techniques: Incubation, Fluorescence, Produced, Transfection, Control, Derivative Assay, Microscopy