Atorvastatin treatment results in the dephosphorylation and inactivation of MYC. (A) Analysis of levels of MYC protein expression and phosphorylation. Levels of MYC and MYC phosphorylated at Ser62 and Thr58 (p-MYC) measured by Western analysis. (B) Temporal analysis of MYC phosphorylation. MYC-induced lymphoma cell lines were treated with atorvastatin and analyzed at indicated times for phosphorylation of ERK1/2, phosphorylation of MYC, and cleavage of caspase 3 by FACS. (C) Treatment of human Burkitt lymphoma cell line ST486 with atorvastatin. Levels of pERK1/2, pMYC, and cleaved caspase 3 were analyzed by FACS following treatment with 10 μM atorvastatin (AT), 100 μM mevalonate (Mev), or 10 μM atorvastatin plus 100 μM mevalonate (AT + Mev). (D) Temporal analysis of MYC phosphorylation. MYC-expressing tumor cells (0.3 × 10⁶) were treated with atorvastatin (10 μM) for different lengths of time and analyzed for changes in MYC phosphorylation and expression of ODC, a known MYC transcriptional target. This image was cropped so that the 0- and 6-hour time points were next to each other. (E) Quantitative real-time PCR analysis of cDNA from MYC lymphoma cells where MYC was expressed (MYC ON) or MYC was inactivated (MYC OFF) after doxycycline treatment (20 ng/mL) or in the presence of 10 μM AT, 100 μM Mev, or 10 μM AT plus 100 μM Mev. Expression of each gene was normalized to the expression of ubiquitin. Changes in gene expression were measured for each treatment and compared with MYC-expressing nontreated tumor cells. Changes greater than 2-fold are statistically significant, as indicated in blue and orange/red (t test: P < .05-.001).