Fig. 3
Fluorescent resonance energy transfer and colocalization of ILK with RI are assayed. a-d EJ cells were co-transfected with plasmids as described previously, and then followed by photobleaching analysis. Images indicated the 488 and cy3 fluorescence emission intensities in the cells before and after photobleaching. e The FRET efficiency (FRETE) was calculated as the percentage of the 488 fluorescence recovery. The experiments were repeated 3 times. f The Intracellular localizations observation of RI and ILK. EJ cells and 293 cells were co-transfected with plamids pEYFP-N1-ILK and pEGFP-C1-RI. The two Proteins labeled with fluorescent proteins revealed a colocalization under Olympus multifunction microscope. g RI expression and colocalization experiment also were observed under laser scanning confocal microscope. Immunofluorescence staining of RI (red) and ILK (green) in four groups of EJ cells co-transfected with different pairs of plasmids. The nucleus was stained with DAPI (blue). The yellow part demonstrated that ILK colocalized with RI in EJ cells. The expression of RI markedly decreased in ILK-upregulated cells compared with the control group cells (magnification × 200)