International Journal of Stem Cells : eISSN 2005-5447

Fig. 2.

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Fig. 2. Effect of NAM treatment on ROS levels, SIRT1 activity, and mitochondria content. (A) BMSCs (from the 31-year-old donor) at PD 8 were incubated without (−) and with (+) 5 mM NAM for 10 days and then stained with MitoSox, DHR123, and DHE to determine the levels of mitochondrial superoxide, hydroxyl radical, and cytosolic superoxide. Values are plotted relative to the cells in day 0 plates. The significance of the difference between (−) and (+) NAM was determined by ANOVA test (Dunnett’s test) (*p<0.1; **p<0.01) (B) BMSCs were cultured in the presence of NAM and the NAD+/NADH ratio was determined at the indicated time points. The significance of the difference compared to the day 0 sample was determined by ANOVA test (Dunnett’s test) (*p<0.1; **p<0.01). (C) BMSCs incubated with 5 mM NAM or 0.16 nM SRT1720 for 0, 1, or 2 days were collected and lysed. The extracts were subjected to western blotting for acetylated or total p53 or Erk protein (upper panel), or LC3 proteins or β-actin (lower panel). (D) The cells were stained with nonyl acridine orange (NAO) to determine mitochondria content.
International Journal of Stem Cells 2018;11:13-25 https://doi.org/10.15283/ijsc18033
© 2018 International Journal of Stem Cells