Effect of NMNAT3v1 or FKSG76 transfection on cellular or mitochondrial NAD and membrane potential.

(A) Whole cellular NAD content in control, NMNAT3v1- and FKSG76-transfected cells. Basal NAD content was 12.6±2 nmol/mg prot. (B) Western blotting evaluation of poly(ADP-ribose) (PAR) formation in HEK cells under control conditions or after transfection of FKSG76 and/or PARP1-cd. Tubulin is shown as loading control. (C) Densitometric evaluation of PAR formation shown in (B). (D) Immunocytochemical localization of PAR in HEK cells under control conditions or after transfection of FKSG76 and/or mitoPARP1cd. (E) Effect of FKSG76 transfection on mitochondrial membrane potential. (F) Representative experiment of the effect of FKSG76 transfection on oxygen consumption. The arrow indicates the time when cells were added to the respiration buffer. (G) Oxygen consumption rate in control or FKSG76-transfected cells. Columns represent the mean ± SEM of 4 (A), 5 (C) and 3 (G) experiments. Western blotting and immunocytochemistry are representative of 5 and 2 experiments, respectively.* p<0.05; ** p<0.01; *** p<0.001 vs control (Student's t test).