At the end of treatment, the cells for the AFM studies were prepared. If necessary, the AT1-receptor blocker candesartan (Astra-Zeneca, Mndal, Sweden) (10 -5 M), and p38 MAP kinase inhibitor SB-203580 (Calbiochem, EMD Biosciences, San Diego, CA) (1 ) was added to the cells in separate experiments, 3 in before ANG II treatment. For the Raf Inhibitors permeability t measurement Carefully washed and the cells were incubated with 1% OsO 4 for 1 h, in ethanol dehydrated ssert, Found with uranyl acetate 1% in ethanol 50% Treated rbt 1 h, and in TAAB 812 (TAAB Laboratory Technology) are embedded. Ultrathin sections were cut from the membrane, with a Leica UCT ultramicrotome (Leica Microsystems, Wetzlar, Germany).

Digital images were recorded on a Hitachi 71 electron microscope (Hitachi, Japan), brightness and contrast if necessary using Adobe Photoshop CS3 (San Jose, CA) set. Acquired for morphometric analysis of images at random, were used to magniations 3 0 7.5 7.5 m image. All experiments were repeated three times. Visualization of endothelial cells endothelial cells by atomic force microscopy were grown on gelatin- EUR glass slides hunter who coated with fibronectin and connce achieved if the complete medium was replaced with medium containing 5% FBS. The cells were then treated with various doses of ANG II (Sigma) or VEGF (R & D Systems) or vehicle. For atomic force Decitabine microscopy (AFM) cells studies were ed by dehydration with increasing ethanol categories. Apical membrane surface Was scanned surface. AFM data were analyzed with scanning software derived from the probe image processor (SPIP, Image Metrology, H Rsholm, D Nemark). Single-cell surface Chen (8 8 m) were scanned and each experimental group experience was at least three to four times (fins) are repeated. On each plate were at least three different cells were examined and two to four samples.

The results show that the average of all the sampled areas. The AFM was originally developed by the University of t Twente (Enschede, The Netherlands) (18) is mounted. In this microscope was the scanning piezo-metric Not directly overhang instead of the sample. calculated Fick’s first law of diffusion: PJ  AC, where P is the result, Herk mmlichen Objekttr ger coefient of glass on a Zeiss permeability t mounted so, the gel substance ste J, A is the effective surface of the Axiophot (Zeiss, Oberkochen, Germany) inverted microscope has endothelialized ter, and C is the concentration used for smooth muscle localization by difference and the choice of cells. Ed strips were on the monolayer and B (4). The Ver Change in the concentration of FITC-dextran 40 in the upper chambers may need during the measurements were negligible Ssigbar. Glass slides hunter and illustrated with Ratschl Gene from silicon nitride (Park Scientific, Sunnyvale, CA) with a radius of curvature of nominally 20 nm, or ultra-high levee Aspektverh Ratio silicon tips (radius 10 nm). AJP-Cell Physiol intermittent 10.2 . 138.2011  .physiology of 6 M Downloaded March, 2012 Page 3 ANG II, PV-1, caveolae C269 tent contact mode was ultra micro levers or type levers used a few tenths of kHz below its resonance frequency (about 120 kHz and 16 respectively). Reagents in controlled The contact mode, Applied Biosystems, Foster City, CA) was a relative quantification using the Bio-Rad CFX Manager software v 1.7. In each experiment, three points were examined in parallel imaging, we have here 512 512 pixel images in the H He and errors and the number of individual experiments are shown in the picture signal modes. The scanned images are not corrected for the folding top, but all samples of a particular experiment were measured with the same tip. Figure 4 pictures were taken in the Department of Biophysics, Semmelweis University t. Non-contact mode AFM images were acquired with an Asylum Research instrument MFP3D .