Figure 1: Everolimus Inhibits SMC Proliferation and Cell Cycle Progression

(A and B) Primary human coronary artery vascular smooth muscle cells (hSMC) were serum-deprived (−) and treated with vehicle or the indicated doses of everolimus (A) or sirolimus (B) in growth medium. Cell counts are expressed as mean ± SEM (n = 12 samples/group) relative to quiescent hSMC. (A) Significance levels versus quiescent cells (−): growth medium + vehicle, *p < 0.001; growth medium + 0.1 μmol/l everolimus, *p = 0.007; growth medium + 1 μmol/l everolimus, *p = 0.009. Significance levels versus growth medium + vehicle: growth medium + 10 μmol/l everolimus, #p < 0.001; growth medium + 25 μmol/l everolimus, #p < 0.001. (B) Significance levels versus quiescent cells (−): growth medium + vehicle, *p < 0.001; growth medium + 0.1 μmol/l sirolimus, *p = 0.010; growth medium + 1 μmol/l sirolimus, *p = 0.008; growth medium + 10 μmol/l sirolimus, *p = 0.008; growth medium + 25 μmol/l sirolimus, *p = 0.009. (C) Deoxyribonucleic acid histograms showing cell cycle distribution of hSMC after treatment with 10 μmol/l everolimus. (D) Western blotting for phosphorylated (phospho) retinoblastoma protein (RB), total RB, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in cell lysates isolated from hSMC. The autoradiograms shown represent 3 independently performed experiments using different cell preparations. Values are normalized to GAPDH and expressed as mean ± SEM (n = 6 samples/group) relative to quiescent hSMC (*p < 0.001 versus starvation).

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