Figure 1.

1a. The expanded insets of 2-dimensional gels reveal the induction of glutaminase (GLS; highlighted by white circles) by Myc in P493-6 B cells. For each condition, 350μg of mitochondrial protein lysate was resolved on 18 cm immobilized pH gradient (IPG) strips as the first dimension followed by 10% Bis-Tris SDS-PAGE as the second dimension, which is marked by molecular mass markers. Protein spots were visualized by silver staining. Six independent biological experiments were performed for each condition. Table S1 summarizes the identity of the spots with the same numbering system as depicted in the figure.

1b. Immunoblot with anti-GLS antibody of a 1-dimensional SDS-PAGE gel of mitochondrial proteins (20 μg/lane) validates the induction of GLS by Myc discovered in Figure 1a. TFAM represents a control mitochondrial protein.

1c. P493-6 cells were treated with tetracycline (Tet) for different lengths of time to inhibit Myc expression or were treated first with tetracycline for 48h and then washed (Wash) to remove tetracycline with the times after wash out indicated. Cells were then harvested for immunoblot assay for GLS or c-Myc. Anti-tubulin antibody and anti-TFAM were used for loading controls.

1d. Human CB33 lymphoblastoid cells, CB33-Myc cells, and PC3 cells transfected with siRNA against c-Myc (siMYC) or control siRNA (siCont) were used for immunoblot assays. Experiments were replicated with similar results.