【文獻標題】Effects of RNA interference–mediated E‐selectin gene silencing on cell adhesionmolecule expression and cell‐cell adhesion in vascular endothelial cells in mice with immunologic contact urticaria
Fourteen days after the model establishment, the endothelia cells were collected according to the aforementioned procedure. The VECs were fixed in 4% formalin for 24 hours, dehydrated by ethanol of grades 80%, 90%, and 100% and N‐butanol, and embedded in paraffin in a paraffin box at 60°C. After cutting the cells into 5‐μm serial sections, cell samples were allowed to stand at 45°C,collected, and dried at 60°C for 1 hour, followed by dewaxing by xylene. After dehydration, the sections were immersed in 3% H2O2 for 10 minutes and rinsed under distilled water. After antigen repair at high pressure for 90 seconds, the sections were cooled down to room temperature. Next, the sections were washed with PBS 3 times for 3 minutes each. Samples were blocked in 5% bovine serum albumin, incubated at 37°C for 30 minutes, and then added with primary antibody rabbit anti‐SELE (100 μL, 1:1000 to 1:4000, ab93952; Abcam,Cambridge, MA) for incubation at 4°C overnight. After washing the vascular endothelium samples with PBS, the samples were incubated with biotinylated goat anti‐rabbit secondary antibody (1:100, HY90046; Shanghai Hengyuan Biological Technology Co, Ltd, Shanghai, China) at 37°C for 30 minutes.
