Figure: siRNA-mediated suppression of target gene expression in Schwann cells. A, Detergent extracts of siNeg- or siGly1-transfected Schwann cells were digested with heparitinase and subjected to immunoblot analysis with anti-glypican-1 antibodies (top); aliquots of undigested extracts were immunoblotted with anti-actin antibodies (bottom) to verify equal sample loading. B, Cell surface expression of glypican-1 was assessed by immunofluorescent staining of transfected cells 48 h after transfection using anti-glypican-1 antibodies (green) and DAPI (4',6'-diamidino-2-phenylindole) to stain nuclei (red). C, Schwann cells were transfected with siNeg or si alpha4(V), and conditioned media and cell lysates were harvested 48 h later (left) or at the indicated times after transfection (right); aliquots of medium (top) or cell lysates (bottom) were subjected to immunoblot analysis and stained with anti-alpha4(V) collagen (top) or anti-actin (bottom) antibodies.