Friday, January 12, 2007

Protocols for Delivering siRNA to Neurons

Here's a link to the protocol: Transfecting Schwann Cells with i-Fect and below are related data.
Figure:si RNA-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 wereimmunoblotted 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 4(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- 4(V) collagen (top) or anti- -actin (b ottom) antibodies.


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