Extended Data Fig. 3: Heightended NADase activity in M1 macrophages is CD38 dependent and PARP1 independent.
From: Senescent cells promote tissue NAD+ decline during ageing via the activation of CD38+ macrophages
a, Flow cytometry results comparing CD38 surface staining in naive (M0) WT and Cd38 KO BMDMs or BMDMs treated with IL-4 (M2) and LPS (M1) for 16 hours. b-c, NADase activity measured with non-cell permeable εNAD in intact M0, M2, and M1 WT and Cd38 KO BMDMs activated for 16 hours relative to cell number (B) and protein content (C). d, mRNA levels of Cd157 in M0 and M1 WT and Cd38 KO BMDMs treated for 16 hours. e, LC-MS quantification of NR in M0 and M1 WT and Cd38 KO BMDMs treated for 16 hours. f, mRNA levels of anti-oxidant genes in WT and Cd38 KO BMDMs treated with IL-4 (M2) and LPS (M1) for the indicated intervals. g, Western analysis of PARP activity (PARylation) and DNA damage (γH2AX) in WT and Cd38 KO BMDMs treated with IL-4 (M2) and LPS (M1) for the indicated intervals compared to WT MO macrophage treated with 1 mM H2O2 for 10 minutes. h, Western analysis of PARP activity (PARylation) and DNA damage (γH2AX) in WT and Cd38 KO BMDMs treated with IL-4 (M2) and LPS (M1) for 8 hours prior to treatment with 1 mM H2O2 for 10 minutes. Data show the mean ± SEM. (n= at least 3 independent experiments). Statistical significance indicated as *P<0.05, **P<0.01, and ***P<0.001; two-sided Student’s t-test. Unless noted with a bar, statistical comparisons are relative to the appropriate MO WT or Cd38 KO sample of the same genotype.
