Figure 1. Differential kinetics of HSC... | Rockefeller University Press

Figure 1.

$Figure 1. Differential kinetics of HSC contribution to blood cell lineages. (A–E) Adult Pdzk1ip1-CreER R26tdTomato reporter mice were given a single dose of tamoxifen, and Tom+ cells were analyzed in the PB of the same continuously monitored animals (A) or in the BM of sacrificed individual mice (B–E). (A) The fraction of Tom+ cells among the indicated cell types in the PB. Shown are values from individual animals (n = 8, pooled from two independent experiments); bars represent median. The significance of differences between granulocytes and platelets as determined by Mann-Whitney test is indicated: **, P < 0.01; ***, P < 0.001; ns, not significant. (B) The expression of Tom versus c-Kit in the total BM cells of a CreER− control or CreER+ reporter mice at the indicated intervals after HSC labeling. Shown are staining plots of individual mice representative of two to four animals per endpoint. (C and D) The phenotype of Tom+ cells in the BM at the indicated time points after HSC labeling. Total BM cells from individual reporter mice (5–6 × 106 cells/animal) were stained with a panel of antibodies to different lineages and progenitor types. All Tom+ cells and 5 × 104 total cells from each of eight samples (four time points, two animals per time point) were concatenated and analyzed using the viSNE algorithm. Each cell was plotted using two resulting tSNE parameters (arbitrary units). (C) The viSNE plot of total cells. Left panels shows major cell types indicated by color (granulocyte, blue; monocyte, light blue; B cell, yellow; T/NK, orange; pDC, brown; LSK, pink; MkP, purple; myeloid, turquoise; erythroid, green); right panels highlight Tom+ cells (red) from each time point. Tom− are indicated in gray. (D) The viSNE plot of gated Tom+ cells. Left panels show major cell types indicated by color (granulocyte, blue; monocyte, light blue; B cell, yellow; pDC, orange; LSK, pink; MkP, purple; myeloid, turquoise; erythroid, green); right panels show cells from each time point highlighted in red. (E) Contour plot overlays of gated Tom+ cells (red) versus all acquired cells (gray) in representative individual samples. Left panels show total cells highlighting c-Kit+ progenitors and CD11b+ myeloid cells. Right panels show gated CD11b− Ter119− cells highlighting B220+ Ly6c+ pDC and B220+ Ly6c− B cells.$

Differential kinetics of HSC contribution to blood cell lineages. (A–E) Adult Pdzk1ip1-CreER R26^tdTomato reporter mice were given a single dose of tamoxifen, and Tom⁺ cells were analyzed in the PB of the same continuously monitored animals (A) or in the BM of sacrificed individual mice (B–E). (A) The fraction of Tom⁺ cells among the indicated cell types in the PB. Shown are values from individual animals (n = 8, pooled from two independent experiments); bars represent median. The significance of differences between granulocytes and platelets as determined by Mann-Whitney test is indicated: **, P < 0.01; ***, P < 0.001; ns, not significant. (B) The expression of Tom versus c-Kit in the total BM cells of a CreER⁻ control or CreER⁺ reporter mice at the indicated intervals after HSC labeling. Shown are staining plots of individual mice representative of two to four animals per endpoint. (C and D) The phenotype of Tom⁺ cells in the BM at the indicated time points after HSC labeling. Total BM cells from individual reporter mice (5–6 × 10⁶ cells/animal) were stained with a panel of antibodies to different lineages and progenitor types. All Tom⁺ cells and 5 × 10⁴ total cells from each of eight samples (four time points, two animals per time point) were concatenated and analyzed using the viSNE algorithm. Each cell was plotted using two resulting tSNE parameters (arbitrary units). (C) The viSNE plot of total cells. Left panels shows major cell types indicated by color (granulocyte, blue; monocyte, light blue; B cell, yellow; T/NK, orange; pDC, brown; LSK, pink; MkP, purple; myeloid, turquoise; erythroid, green); right panels highlight Tom⁺ cells (red) from each time point. Tom⁻ are indicated in gray. (D) The viSNE plot of gated Tom⁺ cells. Left panels show major cell types indicated by color (granulocyte, blue; monocyte, light blue; B cell, yellow; pDC, orange; LSK, pink; MkP, purple; myeloid, turquoise; erythroid, green); right panels show cells from each time point highlighted in red. (E) Contour plot overlays of gated Tom⁺ cells (red) versus all acquired cells (gray) in representative individual samples. Left panels show total cells highlighting c-Kit⁺ progenitors and CD11b⁺ myeloid cells. Right panels show gated CD11b⁻ Ter119⁻ cells highlighting B220⁺ Ly6c⁺ pDC and B220⁺ Ly6c⁻ B cells.

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