Figure 2.
Optimization of EdU-BrdU dual labeling technique applied to HSPC in-vivo A, Dual-deoxynucleoside labeling with single EdU (1.5 mg/mouse) and single BrdU dose (2 mg/mouse) separated by 2 hours (TI). Bone marrow cell were harvested 0.5 hours after BrdU administration. B, Dual-deoxynucleoside labeling with two doses of EdU (2 x 1.5 mg/mouse) and single dose of BrdU (2 mg/mouse). C, Representative dot-plots of Lin−cKit+ cells and EdU/BrdU fractions frequency in three experimental settings. D, Ratio of S-phase exiting cells (EdU+BrdU−) to S-phase entering cells (EdU−BrdU+) in three experimental settings determined in LSK cells, LS−K cells and their CMP, GMP and MEP subpopulations. Four to six mice were analyzed in each of the experimental settings. The experimentally obtained ratios were tested by Student’s t-test (one sample, two-tailed) to the expected theoretical ratio of 4.0 or 3.0 (see text). The ratios were significantly different (P < 0.05) from 4.0 in the left two columns. Results in the table are mean ± SEM.