Extended Data Fig. 7: LPS promotes tissue NAD decline via CD38.

a, Representative gating for the splenic leukocyte populations quantified in Fig. 7c, d and Extended Data Fig. 7a. Left six panels show gating for identification of B cells and different myeloid cells, as indicated, as well as gating for CD38-positive cells in all populations. Right six panels show gating for T cell subsets, as indicated. Red arrows indicate sequential gating, pointing from parent plots towards child plots. b, Quantification of immune cell populations and CD38+ immune cells in the spleen of 3 month-old WT male mice i.p. injected with PBS or LPS for 4 weeks, and analyzed by flow cytometry. (PBS n=10 mice/group, LPS n=9 mice/group) c, Western analysis of CD38, CD157, CD68, and NAMPT in eWAT of 3 month-old WT male mice injected with PBS or LPS for 4 weeks and Image J quantification of CD38 protein levels relative to NAMPT. Each lane represents one mouse (PBS n=4 mice/group, LPS n=5 mice/group) d, mRNA levels of NAD consuming enzymes in eWAT from 3 month-old WT male mice injected with PBS or LPS for 4 weeks. (PBS n=4 mice/group, LPS n=5 mice/group) e, mRNA levels of the indicated genes in whole eWAT from 4 month-old WT and Cd38 KO male mice injected with PBS or LPS for 12 hours. (n=10 mice/group) f, Western analysis of eWAT from 4 month-old WT and Cd38 KO male mice injected with PBS or LPS for 12 hours (n=3 mice/group). g, LC-MS quantification of NAD-related metabolites in eWAT from 4 month-old WT and Cd38 KO male mice IP injected with PBS or LPS for 12 hours. (n=10 mice/group) h, LC-MS quantification of NAD-related metabolites in liver from 4 month-old WT and Cd38 KO male mice IP injected with PBS or LPS for 12 hours. (n=10 mice/group) Data from individual mice are shown for in vivo experiments. Data show the mean ± SEM. Statistical significance indicated as *P<0.05, **P<0.01, and ***P<0.001; two-sided Student’s t-test except for 7 g and 7 h one-tailed t-test was used.

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