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4-OHT time release pellets (5?mg; Innovative Research of America) were inserted subcutaneously at age of 18?weeks to reach steady-state blood levels of 1?ng ml-1 4-OHT for 60 consecutive days. Antibodies used for IHC include anti-Ki67 (Dako), anti-53BP1 (Bethyl Labs), anti-��H2AX (Bethyl Labs), and anti-Cleaved Caspase 3 (Cell Signaling). For FACS analysis, cells were immunostained with CD4, CD8, and CD3 antibodies (eBioscience) and analyzed on a BD FACSCanto II (BD Biosciences). For metaphase preparation, metaphases were obtained from colcemid-treated cells incubated in 105?mM KCl hypotonic buffer for 15?min before fixation in 3:1 methanol�Cacetic acid. Spectral karyotyping (SKY) was done using the SkyPaint Kit and SkyView analytical software (Applied Spectral Imaging) according to manufacturer's CH5424802 supplier protocols. For telomere-FISH (fluorescent in?situ hybridization), metaphase spreads were applied VX-770 solubility dmso with telomere-specific T2AG3-FITC PNA (peptide nucleic acid) probes and centromere-specific Cent-Cy3 or Cent-pacific blue PNA probes, and counter-stained with DAPI (for?microscopy) or TOTO3 (for laser scanning). Laser scanning cytometry quantification was performed with an iCys Research Imaging Cytometer (Compucyte) as described earlier (Jaskelioff et?al., 2011?and?Sahin et?al., 2011). Telomere Co-FISH staining was performed as previously described (Potts and Yu, 2007). Details are described in Extended Experimental Procedures. RT-qPCR primers are described in Table S4. Mitochondrial DNA content was measured by the relative levels of CoxI versus beta globin by qPCR. CoxI and beta globin primers are described in Table S4. Antibodies used for Western blotting are anti-phospho p53 (ser15, Cell Signaling), anti-p21 (Santa Cruz Biotechnology), anti-Vinculin (Sigma), and anti-Mre11 (BD Biosciences). Lentiviral based shRNA constructs targeting mouse PGC-1�� and SOD2 were ordered from The Dana-Farber/Harvard Cancer Center DNA Resource Core. Verified shRNA sequences were cloned into inducible lentiviral constructs PLKO-TRC-901 (IPTG inducible; Broad Institute) and PLKO-Tet (Doxycycline inducible; Novartis). Eight hundred Ribonucleotide reductase ninety-one upregulated and 1,345 downregulated genes with 1.5-fold change and p?