Transfection of adherent 3T3-L1 adipocytes with siRNA does not decrease expression of targeted proteins.

<p>Forty-eight hours post-transfection, transfection efficiency was assayed as uptake of fluorescent-labeled siRNA and the siRNA-mediated effect on targeted protein expression. (A) Brightfield and fluorescent (rhodamine filter) images of the transfected cells were taken with a 40X objective. (B) Knockdown of lamin A/C and PPARγ was assayed via western blot analysis post-transfection with either the RISC-free control siRNA (control), nontargeting siRNA (NT), or siRNA targeted to the indicated gene (target). Thirty-five µg of protein was loaded in each lane and separated by SDS-PAGE. Equal loading of each lane was determined using β-actin expression. The experiment was carried out twice independently.