Oxidative Medicine and Cellular Longevity / 2018 / Article / Fig 8

Research Article

Parathyroid Hormone Causes Endothelial Dysfunction by Inducing Mitochondrial ROS and Specific Oxidative Signal Transduction Modifications

Figure 8

Characterization of angiogenic phenotype in response to PTH (a–e). BAECs were plated on Matrigel matrix, and tubular formations were evaluated 24 h after PTH (0.1 nM) treatment and compared to control. Pictures of tubular formations are obtained using an optical microscope; the images are representative of results from three independent experiments. The scale bar is about 20 μm (a). The angiogenic response was quantized as the number of branch points, and the data are reported as mean ± SD ( vs. CTRL) (b). From BAECs, after 24 h of PTH treatment, mRNA was isolated; the transcription levels of VEGF were evaluated by real-time PCR and expressed as fold increase vs. control ( vs. CTRL) (c). VEGF receptor was immunoprecipitated from cells treated with PTH for 24 h. The levels of VEGF receptor oxidation were determined by OxyBlot Protein Detection Kit as described in Materials and Methods and shown as levels of 2,4-dinitrophenylhydrazine (DNP) bound to the receptor. CTRL− represents an immunoprecipitation using the secondary antibody to test signal specificity. CTRL+ represents a whole cell lysate. The images are representative of results from three independent experiments (d). Densitometric analysis of oxidized VEGFR2 is shown in the bar graph as mean ± SD () of fold change vs. control (e). The image is the mean of three independent experiments.