Fig. 7.

Effect of NAMPT Knock Down and NR Rescue On GLO1 Protein Expression and Activity in Human Immortalized Myotubes

Human Immortalized Myotubes transfected with NAMPT siRNA for 48 h, or siRNA for 48 h with 0.5 mM nicotinamide riboside (NR) treatment for the final 24 h (NAMPT siRNA + 0.5 mM NR). NAMPT siRNA depleted myotubes of NAMPT protein (A), and transcripts (B) which were not rescued by NR. The effects of NAMPT KD and NR rescue on GLO1 protein abundance was determined via Western blot (C) and ELISA (D). GLO1 specific activity (F) was determined by making GLO1 activity (E) relative to GLO1 abundance in each sample which was determined via ELISA. A representative Western blot for GLO1, NAMPT and total protein used for normalizing signal quantification of target proteins are displayed in panel G. Data are presented as mean ± SEM with individual data points superimposed. Data were compared via One-way ANOVA using Bonferroni Post Hoc analysis with significance set to p < 0.05.