mouse cd38 proteins  (R&D Systems)

Journal: Nature metabolism

Article Title: CD38 ecto-enzyme in immune cells is induced during aging and regulates NAD + and NMN levels.

doi: 10.1038/s42255-020-00298-z

Figure Lengend Snippet: Fig. 4 | SASP increases CD38 accumulation and decreases NAD+ levels in WAT in vivo. a,b, Twelve-month-old WT mice received daily s.c. injection of CM from CM-SEN or CM-NS mouse pre-adipocytes for 5 d, and tissues were harvested at day 5. a, CD38 activity (n = 7 mice per group) and NAD+ levels (CM-NS, n = 7 mice; CM-SEN, n = 6 mice) were measured in subcutaneous (subq) WAT. b, Relative Cd38 (n = 11 mice per group), Cd45 and F4/80 (CM-NS, n = 14 mice; CM-SEN, n = 12 mice) mRNA expression measured by RT–qPCR in subq WAT. Levels were relative to CM-NS. c–f, Twenty-six-month-old mice were treated with vehicle (ctrl) or 3TC for 2 weeks. c, Schematic of experiment. d, Relative mRNA levels for p16 and SASP components were measured by RT–qPCR in WAT (n = 8 mice per group for all conditions except Pai1 3TC (n = 7)). e, Relative mRNA levels for F4/80 (n = 8 mice per group), Cd38, Parp1 and Sirt1 (n = 5 mice per group) were measured by RT–qPCR in WAT. f, Relative NAD+ levels in WAT (n = 5 mice per group); levels were relative to vehicle-treated mice. g, Immunofluorescence staining of CD38 (red), lamin B1 (green) and IL-6 (yellow) in WAT of young (2-month-old) and old (28-month-old) C57BL/6 mice, representative of n = 4 young mice and n = 6 old mice. Insets show enlarged areas of old tissue, with and without IL-6 staining, to highlight that CD38 can be found near IL-6+lamin B1− cells. Data are the mean ± s.e.m., analysed by an unpaired two-sided t-test. Two statistical outliers were identified in the Cd45 mRNA expression of the CM-SEN-treated animals (with values >16) using the appropriate test in GraphPad Prism 6. These samples were excluded from all PCR analyses for this experiment. Data were also analysed by non-parametric Mann–Whitney test using the full dataset (including the identified statistical outliers); Cd45 P value = 0.002.

Article Snippet: In brief, the lymph nodes of six UniRats immunized with recombinant murine CD38 protein (R&D Systems) were isolated, washed and cell pellets frozen.

Techniques: In Vivo, Injection, Activity Assay, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, MANN-WHITNEY

Journal: Nature metabolism

Article Title: CD38 ecto-enzyme in immune cells is induced during aging and regulates NAD + and NMN levels.

doi: 10.1038/s42255-020-00298-z

Figure Lengend Snippet: Fig. 9 | Schematic of the role of cellular senescence and sterile inflammation in the regulation of CD38 and NMN degradation. In young mice, levels of CD38+ inflammatory cells and senescent cells in tissues are lower than in aged mice. During aging, there is an increase in senescent cells that, in part, through their SASP, promotes accumulation of CD38+ immune cells. The ecto-enzymatic activity of CD38 in immune cells degrades NMN extracellularly, preventing the NMN-induced NAD+ boosting in other cells in the tissue.

Article Snippet: In brief, the lymph nodes of six UniRats immunized with recombinant murine CD38 protein (R&D Systems) were isolated, washed and cell pellets frozen.

Techniques: Sterility, Activity Assay