Effect of increasing concentrations of simvastatin on PCSK9 and LDLR expression in HepG2 cells. HepG2 cells were grown in DMEM + 10% FBS and supplemented with increasing concentrations of simvastatin for 24 hours at 37°C. 50 μg of total cell lysates (A and C) and 100 μl cell media (B) were analyzed by immunoblotting (IB), or immunoprecipitation (IP) followed by IB, respectively, as described in Methods. A and B were IB with anti-hPCSK9 Ab and C with anti-hLDLR Ab. The protein signals were quantified by densitometry using Syngene Chemigenius 2XE imager and Gene Tools software. All values were made relative to values from untreated cells set as 1 and presented as mean ± SEM (n = 3). * Indicates significant differences from untreated control (p < 0.05) using student's T-test.