Extended Data Fig. 2: Analysis of the role of the NAM-salvage pathway and sirtuins in macrophage activation and polarization.
From: Senescent cells promote tissue NAD+ decline during ageing via the activation of CD38+ macrophages
a, NAD levels quantified by LC-MS in M0, M1 and M2 BMDMs pre-treated or not with 50 nM FK866 and NR for 6 hours prior to stimulation with LPS for an additional 6 hours or IL-4 for 16 hours. b, mRNA levels of M2 genes in BMDMs pre-treated or not with FK866 and NR for 6 hours prior to stimulation with IL-4 for 16 hours. All statistical comparisons are relative to M2 + FK866. c, mRNA levels of M1 genes in BMDMs pre-treated or not with FK866 and NR for 6 hours prior to stimulation with LPS for 6 hours. All statistical comparisons are relative to M1 + FK866. d, Western analysis of Nampt Fl/Fl CreER and Nampt Fl/Fl BMDMs treated with 1 ug/ml tamoxifen. e, mRNA levels of M2 genes in Nampt Fl/Fl CreER and Nampt Fl/Fl BMDMs treated with IL-4 for 16 hours. f, mRNA levels of M1 genes in Nampt Fl/Fl CreER and Nampt Fl/Fl BMDMs treated with LPS for 6 hours. g, mRNA levels of M2 genes in WT BMDMs pretreated with the indicated concentration of the sirtuin inhibitors Ex527 and AGK2 for 30 minutes prior to stimulation with IL-4 for 16 hours. All statistical comparisons are relative to M2. h, mRNA levels of M1 genes in WT BMDMs pretreated with the indicated concentration of the sirtuin inhibitors Ex527 and AGK2 for 30 minutes prior to stimulation with LPS for 6 hours. All statistical comparisons are relative to M1. Data show the mean ± SEM. (n=3 independent experiments). Statistical significance defined as *P<0.05, **P<0.01, and ***P<0.001; two-sided Student’s t-test.
