Figure 5

Decrease of uPAR-mediated adhesion to vitronectin by knockdown of MRJ and/or HSP70 in HEK293-uPAR and HEK293T cells. (A) Adhesion assay. The cells were transfected with psiSC, or psiMRJ, or psiHSP70 or psiMRJ plus psiHSP70 and grown on 96-well plates pre-coated with vitronectin (1 μg/ml) and stained with Giemsa and then quantified using Safire Fluorescence Absorbance and Luminescence Reader (TECAN). *P < 0.05: significantly different from the control. The experiments were repeated twice and performed in triplicate. (B) Reduction of uPAR on the cell surface by knockdown of MRJ/HSP70 using flow cytometry in HEK293-uPAR cells. The cells were transfected with psiSC, or psiMRJ, or psiHSP70, or psiMRJ plus psiHSP70. Forty-eight hours after transfection, cells were harvested and labeled with anti-human uPAR monoclonal antibody for 30 min. After labeling, the cells were immediately analysed by flow cytometry in APC channel. Suitable negative isotype controls were used to rule out the background fluorescence.