Figure 5.

siRNA-mediated LSAMP knockdown enhances aortic SMC proliferation. Human aortic SMCs were transfected with siRNA targeting either LSAMP or no known gene (control), and assayed after 72 hours in serum-free medium. A, Membrane fractions of SMCs (50 μg) and whole mouse brain (20 μg, as a positive control) were subjected to SDS-polyacrylamide gel electrophoresis and immunoblotting (IB) with mouse monoclonal anti-LSAMP or non-immune control IgG. The LSAMP blot was stripped and re-probed for tubulin (as a loading control). Shown is an IB from a single experiment, representative of two performed in duplicate. B, LSAMP expression was measured on total RNA by real-time RT-PCR in triplicate, and normalized to GAPDH expression. *, P < 0.05. C, Quiescent SMCs were fed serum-free medium lacking (basal) or containing 5% fetal bovine serum (FBS) for 24 hours, and [³H]thymidine was added to the medium during the last 4 hours. Incorporation of [³H]thymidine was determined. Shown are the results of a single experiment performed in triplicate, representative of two independent experiments. *, P <0.001 compared with basal; #, P < 0.01 compared with stimulated control SMCs.