Research Paper Volume 8, Issue 1 pp 34—47

Sirt6 regulates dendritic cell differentiation, maturation, and function

Figure 3. In vitro generated Sirt6KO BMDCs show increased endocytic activity and impaired allostimulatory capacity. (A, B) WT and Sirt6KO BMDCs were harvested at day 7 and incubated with dextran-FITC for 30 min at 37°C or at 4°C. Thereafter, cells were stained for CD11c and finally analyzed by flow cytometry. (A) Gating was done on CD11c+ cells; one representative experiment out of six is presented. (B) Dextran-FITC+/CD11c+ Sirt6KO BMDCs were enumerated and their frequency was normalized to that of dextran-FITC+/CD11c+ WT BMDCs. Results are means ± SEM of six separate experiments, n=6 for each genotype. (C) purified allogeneic (BALB-c) CD4+ splenocytes (responders) were stained with CFSE and incubated with 5 μg/ml phythoemagglutinin (PHA) or with sorted, WT or Sirt6KO, CD11c+MHCII+ BMDCs at the indicated S:R ratios. Proliferation of alive (propidium-iodide negative) CD3+CD4+CD11c cells was evaluated by carboxyfluorescein succinimidyl ester (CFSE) dilution after a 5-day incubation. One representative experiment out of three is presented, n=4 for each genotype.