Figure 2
Statin effect in primary human myotubes on mevalonate and cholesterol dependent gene expression level. (A) HMGCR transcript variants amplified for exon 1–20 in primary human myotubes (cell population C11) and liver HepG2 cells. Primary human muscle cells and HepG2 cells share the identical HMGCR transcripts. Unspecific bands in lane 6 were sequenced. [1, Exon1–5; 2, Exon 3-8; 3, Exon 6–11; 4, Exon 9-14; 5, Exon 12-17; 6, Exon 15-20; 7, Exon 1-20; full-length gel was cropped without losing any relevant information, original picture scan in Supplementary Fig. S7] (B) Mevalonate levels in primary human myoblasts after 72 h statin treatment at different concentrations in delipidated medium. Mevalonate levels correlate to cholesterol levels. Data were normalized to total protein level and relativized to DMSO treated samples. (n = 4; cell populations C1, C9, C11, C16) (C) Relative cholesterol levels were determined under delipidated medium conditions in primary human myoblasts after 72 h statin treatment at different concentrations. (error/bars represent mean with SD; n = 3; cell populations C4, C9, C17) (D) Relative mRNA expression of HMGCR, HMGCS, and LDLR were examined in primary human myoblasts undergoing differentiation under statin treatment (5 µM) for 72 h-96 h. The results were normalized to two reference genes and calculated relatively to DMSO-treated samples. (error/bars represent mean with SEM; n = 6; cell populations C2-C4, C11-C13). For cell populations used see Supplementary Table S1. [UT, untreated (black); Sim, simvastatin (red); Rosu, rosuvastatin (blue); a = 50 µM, b = 5 µM, c = 1 µM; HMGCR, 3-hydroxy-3-methylglutaryl-CoA reductase; HMGCS, 3-hydroxy-3-methylglutaryl-CoA synthase 1; LDLR, low density lipoprotein receptor].
