Research Paper Volume 12, Issue 24 pp 25581—25598

Figure 3. Effects of different concentrations of JMF3086 on HMG-CoA reductase expression and enzymatic activity in SH-SY5Y cells stably transfected with LRRK2-G2019S. (A) Western blot of HMG-CoA reductase protein expression after treatment with different concentrations of JMF3086. (B) Quantification of drug effects on the ratio of HMG-CoA reductase to GAPDH. (C) Quantification of HMG-CoA reductase enzymatic activity after treatment with different concentrations of JMF3086. Experiments were repeated in triplicate, and the ratio was compared to SH-SY5Y control cells without transgenes and drug treatment. The relative HMG-CoA reductase enzymatic activity of LRRK2-WT, LRRK2-G2019S without treatment, or after treatment with DMSO solvent, 0.05 μM, 0.1 μM, 0.5 μM, and 1.0 μM JMF3086 was 1.09±0.03, 1.10±0.02, 1.04±0.02, 0.92±0.04, 0.88±0.08, 0.78±0.06, and 0.74±0.03, respectively. P=0.04 for LRRK2-G2019S vs. LRRK2-G2019S with 0.5 μM or 1.0 μM JMF3086, one-way ANOVA. All neurons were treated for 48 hours and then lysed. Equal amounts of protein lysate were subjected to SDS-PAGE and the proteins analyzed by Western blotting. Immunoblots were probed with the indicated antibodies. Data represent mean ± SEM. *P<0.05, **P<0.01.