Rpl13a

Flow cytometry and quantitative Western analysis were performed as previously described.^{20 (link)} Antibodies used in flow cytometry were the following: Tuj1 (Covance, 1:2500), MAP2 (BD Biosciences, Franklin Lakes, NJ; 1:40), glial fibrillary acidic protein (GFAP; Millipore, 1:20), Tra1–81 (Millipore, 1:500), and SSEA3 (Millipore, 1:500). Secondary antibodies were antimouse Alexa 488 (1:1,000), antirat Alexa 488 (1:1,000), or antirabbit Alexa 647 (1:1,000). The following antibodies were used in quantitative Western analysis: frataxin (MitoSciences, Eugene, OR; 1:1,000), RPL13a (Cell Signaling Technology, Danvers, MA; 1:2,000), and RNA polymerase II (Millipore; 1:2,000). Antibodies against acetylated residues of histone H3 and H4 have been described.^{22 (link)} The following secondary antibodies were all obtained from LI-COR Biosciences (Lincoln, NE) and used at the same dilution (1:5,000): antimouse IR680, antimouse IR800, antirabbit IR680, and antirabbit IR800.

To generate enough material for Western blotting, cells were seeded at 175,000/15 cm plates (for samples collected up to 6 days after treatment) or at a density of 300 cells/well in 6-well plates (for samples at 9 days after treatment). Cell extracts were prepared in RIPA buffer and analyzed by 10% SDS-PAGE. Western blots were probed with anti-PARP (1:750; cat. no. 9542, Cell Signaling Technology); anti-γ-tubulin (1:1,000; cat. no. T-5192, Sigma-Aldrich); anti-eEF1A, eEF2 or RPL13a (rabbit, 1:1,000; Cell Signaling Technology); anti-cyclin D1 (rabbit, 1:250–500; cat. no. SC-753, Santa Cruz Biotechnology® Inc., Dallas, TX); anti-nucleolin (mouse, 1:5,000; cat. no. SC8031, Santa Cruz Biotechnology); anti-Cdk1 (1:1,000; cat. no. 9116, Cell Signaling Technology), anti-phospho-Y15 Cdk1 (1:1,000; cat. no. 9111, Cell Signaling Technology); anti-p21 (1:500; Cell Signaling Technology); anti-γ-H2Ax (1:250; Abcam); HRP-conjugated secondary antibodies and SuperSignal West Pico Solutions (Thermo Fisher Scientific Inc., Waltham, MA).