Supplementary Fig 5.

Uptake of simvastatin by undifferentiated and differentiated C2C12 cells. The uptake was determined after incubation of the cells with 1 μmol/L of simvastatin lactone (SVL) or simvastatin hydroxy acid (SVA) in Dulbecco's modified eagle medium of different pH levels (6.8–7.8) at 37°C for 6 hours. Results are expressed as nanomoles per milligram of protein ± standard deviation, (n = 5). Data were analyzed by 2-way analysis of variance followed by Bonferroni's post hoc test. (A) Intracellular concentration of simvastatin acid and lactone recovered from undifferentiated cells after treatment with 1 μmol/L SVL. a = SVL, pH 6.8 vs SVA, pH 6.8; SVL, pH 7.4 vs SVA, pH 7.4 (P < 0.001); b = SVL, pH 6.8 vs SVA, pH 6.8; SVL, pH 7.4 vs SVA, pH 7.4; SVL, pH 7.8 vs SVA, pH 7.8 (P < 0.001); c = SVL, pH 6.8 vs SVA, pH 6.8; SVL, pH 7.4 vs SVA, pH 7.4 (P < 0.001); SVL, pH 7.8 vs SVA, pH 7.8 (P < 0.01). (B) Intracellular concentration of simvastatin acid and lactone recovered from differentiated cells after treatment with 1 μmol/L SVL. a = SVL, pH 6.8 vs SVA, pH 6.8 (P < 0.001). (C) Intracellular concentration of simvastatin acid and lactone recovered from differentiated cells after treatment with 1 μmol/L SVA. a = SVA, pH 6.8 vs SVL, pH 6.8; SVA, pH 7.4 vs SVL, pH 7.4 (P < 0.001); b = SVA, pH 6.8 vs SVL, pH 6.8; SVA, pH 7.4 vs SVL, pH 7.4; SVA, pH 7.8 vs SVL, pH 7.8 (P < 0.001); c = SVA, pH 6.8 vs SVL, pH 6.8; SVA, pH 7.4 vs SVL, pH 7.4 (P < 0.001); SVA, pH 7.8 vs SVL, pH 7.8 (P < 0.01).