Fig. 4

ENO1-silencing modulates ECM-receptor expression. a Using quantitative PCR, mRNA coding for different proteins was investigated in shENO1 CFPAC-1 cells. Values are expressed as relative expression compared to control cells. b Western blot analysis was carried out to investigate alpha 5, beta 1, alpha v, beta 3, and uPAR expression on total lysates of shCTRL and shENO1 cells. c To determine surface expression of integrins, shENO1, or shCTRL cells were incubated with primary antibodies (gray peak) against beta 1, alpha v/beta 3, alpha IIb/beta 3, and or isotype-matched control antibody (empty peak) and analyzed by flow cytometry. d The adhesion ability of shENO1 and shCTRL cells on vitronectin was evaluated in the presence of anti-beta 3 Ab. Adherent cells were fixed with 2% glutaraldehyde in PBS and visualized by staining with crystal violet. For quantitative analysis, cells were treated with 10% acetic acid and elutes were read with a microplate reader at a wavelength of 570 nm. Results are expressed as OD, optical density units. A representative of three independent experiments is shown. Data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 relative to control cells