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. 2013 Oct 14;8(10):e76938. doi: 10.1371/journal.pone.0076938

Figure 6. Effect of exogenous NAD or its precursors on cellular and mitochondrial NAD.

Figure 6

(A) Quantification of NAD in control or FKSG67-transfected HEK cells. The effect of NAD, nicotinamide (Nam), NMN, nicotinamide riboside (NR) or nicotinic acid (NA) on NAD contents of FKSG-transfected cells is shown (NAD and its precursors have been added to the incubating media at 1 mM for 48 hrs). Basal NAD content was 12.6±2 nmol/mg prot. (B) Western blotting evaluation of the effect of NAD, Nam, NMN, NR or nicotinic acid NA (1 mM/48 hrs) on depletion of mitochondrial PAR content induced by FKSG76 co-transfection in mitoPARP1cd-transfected cells. Tubulin is shown as loading control. (C) Densitometric analysis on the experiment shown in (B). (D) Effect of oligomycin (10 µM/30 min) and/or glucose deprivation (30 min) on cellular ATP contents. (E) Effects of exogenous NAD (1 mM/3 hrs) on mitochondrial PAR contents in mitoPARP1cd-transfected cells under control conditions or exposed to oligomycin (10 µM) in the presence or absence of glucose. (F) Densitometric analysis on the experiment shown in (E). Columns represent the mean ± SEM of 3 experiments. Western blotting is representative of 3 (E) and 4 (C) experiments.* p<0.05; ** p<0.01 vs control (Student's t test).