Fig. 8

Systematic comparison of various age-associated changes in immune cell regeneration. A Changes in the production of different types of immune cells between the pre-divergent and end time points in early (n = 9) and delayed (n = 9) aging mice. Each marker indicates one mouse; each horizontal black bar denotes the mean of all the mice in each group. The change is shown on a “symlog” scale with a linear scale ranging between 0 and 100. Bonferroni adjusted Wilcoxon rank-sum test. B Differences in the average change in immune cell production between early aging and delayed aging mice, as shown in A. C A partial least squares regression (PLSR) model was used to determine the factors influencing the population-level B-cell-to-granulocyte ratio at the end of the study. A comparison of the predicted PLSR model outputs and the experimental measurements is shown. D Variable importance of projection (VIP) scores generated by the PLSR model. Generally, a VIP score greater than one means that the input is highly influential on the output. The pre-divergent and end-time-point ratios were determined at the cell population level via FACS analyses. The abundance of each lineage and their changes were determined on the basis of clonal-level measurements. E The final model weights are shown to illustrate the amount of positive or negative influence of each parameter. F Pearson’s correlation (R) between the normalized changes in population-level granulocyte and B-cell abundances in the peripheral blood. The normalized changes were calculated as the differences in abundances between the initial and the end time points divided by the abundances at the initial time point. G Primary mouse HSCs from Cas9-expressing transgenic mice were transduced with lentivirus carrying a mixture of sgRNAs targeting each gene. A non-targeting (NT) sgRNA mixture was used as a control. The abundance of granulocytes (Mac1⁺ Gr1⁺ ) among all living cells after 10 days of co-culturing HSCs with OP9 cells is shown. Two or more biological replicates with a total of 14 or more replicates for each gene were performed. The data are shown as the means ± SEMs. Two-tailed Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001