BrdU-labeling kinetics of the total DC population of different lymphoid organs.
BrdU was administered continually to groups of 4 to 6 mice, and the DCs were isolated at different times from the different pooled lymphoid organs. DCs were surface stained for DC markers, then permeabilized and stained for BrdU incorporated into DNA. DC suspension was then analyzed by flow cytometry, gating for CD11c+MHC class IIhi DCs. Percentages of DCs labeled with BrdU were then determined from profiles similar to those in Figure 1. A parallel group of control mice without BrdU administration provided the DCs for the background control. Results represent pooled data from 3 to 5 separate kinetic experiments. The point near zero time was a 2-hour pulse of BrdU, which should label any DC in cell cycle; later points represent the accumulation of labeled cells from dividing precursors.
