FIGURE 7.

Ectopic expression of LOX and LOXL2 alone reduces E-cadherin expression in RCC10/VHL cells. A, RCC10/VHL cells were transiently transfected with expression plasmids bearing either LOX (pcLOX) or LOXL2 (pcLOXL2) cDNA 72 h prior to harvesting the cells and analyzed by immunoblotting. Equimolar amounts of empty vector (pcDNA) were transfected as a control. B, transient transfection of LOXL2 cDNA, which is tagged with a FLAG tag. Transfection was purposely performed less efficiently than in A, in order to be able to compare negative cells with positively transfected cells. Cells were fixed 48 h after transfection and analyzed by immunofluorescence for E-cadherin, LOXL2-FLAG tag, and DAPI nuclear stain. The arrows indicate adjacent RCC10/VHL cells, which have lost E-cadherin at their site of contact. Green, α-E-cadherin; red, α-FLAG staining; blue, DAPI; bar, 10 μm.