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. Author manuscript; available in PMC: 2015 Nov 4.
Published in final edited form as: Cell Metab. 2014 Nov 4;20(5):840–855. doi: 10.1016/j.cmet.2014.10.005

Fig. 3. PARP1 activation drives SIRT1 depression and the mitochondrial phenotype in CSB deficient cells.

Fig. 3

(A) Representative confocal microscopy images and quantification of WT and CSB deficient cells stained for PARP1 and PAR (n=3, mean ± SEM). (B) Representative immunoblot of PAR, PARP1, PARG1 and UCP2 in WT and CSB deficient cells (n=3, mean ± SEM). (C) Mass spectrometry of nicotinamide adenine dinucleotide (NAD) and NAD-metabolites in WT and CSB deficient cells; NMN: Nicotinamide mononucleotide; NAM: Nicotinamide; (n=6–8, mean ± SEM). (D) Representative immunoblot of sirtuin levels in WT and CSB deficient cells. (E) Immunoblot of protein levels from the cerebellum of WT and Csbm/m mice on various diets. Each lane is a separate mouse. (F) FCCP uncoupled respiration normalized to basal respiration under various 24 hour treatments (n=3–28 separate seahorse experiments, mean ± SEM). (G) FCCP uncoupled respiration normalized to basal respiration 72 hours after SIRT1 siRNA treatment (n=3, mean ± SEM). (H) FCCP uncoupled respiration normalized to basal respiration 72 hours after UCP2 overexpression (n=3, mean ± SEM). (I) Flow cytometry of WT and CSB deficient cells after overexpression of UCP2 and stained with tetramethylrhodamine methyl ester (TMRM) for membrane potential and mitosox for mitochondrial superoxide production (n=3, mean ± SEM).