Figure 1.
Fate mapping and proliferation tracking of perturbed hematopoiesis. (A) How perturbations of hematopoiesis alter contribution of a primitive HSC subset will be studied by fate mapping (red dots and arrow) and proliferation tracking (green shades and arrow). (B) Fgd5ZsGreen:CreERT2/R26LSL-tdRFP mouse model for fate mapping of HSCs. (C-D) RFP labeling of HSPCs isolated from TAM-induced Fgd5ZsGreen:CreERT2/R26LSL-tdRFP mice. Data from Morcos et al.22 (C) Initial labeling frequencies of BM HSPCs 10 days after induction. (D) Labeling of total HSCs (black line) relative to ES HSCs (dotted line, 8-20 mice per time point). (E) R26rtTA/Col1A1H2B-GFP mouse model for proliferation tracking of HSPCs. (F) Representative H2B-GFP histograms of ES HSCs isolated from 5-FU–perturbed and control mice (supplemental Figure 3C-E). (G-M) Fgd5ZsGreen:CreERT2/R26LSL-tdRFP mice were TAM-induced and perturbed with either 2 Gy γ-radiation (n = 4), 5-FU (n = 5) or left untreated (n = 5). Experiment scheme (G), PB lymphocyte count 7 days after irradiation (H), PB reticulocyte numbers 7 days after 5-FU (I), and ratios of relative BM compartment sizes (% cells among total lin− BM cells) between perturbed and control (dotted line) animals (J-K) are shown. (L) Percentages of RFP+ ES HSCs. (M) Fraction of RFP-labeled cells relative to ES HSCs (significant differences between untreated and either 5-FU–perturbed or irradiated mice was calculated, all comparisons between unperturbed and irradiated mice are not significant). (N-O) R26rtTA/Col1A1H2B-GFP animals were doxycycline (DOX)–pulsed and exposed to either 2 Gy (n = 5), 5-FU (n = 5), or left untreated (n = 6). Experiment scheme (N) and numbers of additional divisions in response to perturbation (O) are shown (supplemental Figure 3C-E). (P) Transformation of data shown in panel M to visualize the net effects of myeloablation on label propagation. Mean (dark gray line) and variance (standard deviation [SD], shaded area) of control mice (n = 41 from 8 independent experiments) are shown (supplemental Figure 3F-H). CMP, common myeloid progenitor; Ery, erythrocyte; MkP, megakaryocyte progenitor; Neut, neutrophil; Plt, platelet; PMN, polymorphonuclear cell.

Fate mapping and proliferation tracking of perturbed hematopoiesis. (A) How perturbations of hematopoiesis alter contribution of a primitive HSC subset will be studied by fate mapping (red dots and arrow) and proliferation tracking (green shades and arrow). (B) Fgd5ZsGreen:CreERT2/R26LSL-tdRFP mouse model for fate mapping of HSCs. (C-D) RFP labeling of HSPCs isolated from TAM-induced Fgd5ZsGreen:CreERT2/R26LSL-tdRFP mice. Data from Morcos et al.22 (C) Initial labeling frequencies of BM HSPCs 10 days after induction. (D) Labeling of total HSCs (black line) relative to ES HSCs (dotted line, 8-20 mice per time point). (E) R26rtTA/Col1A1H2B-GFP mouse model for proliferation tracking of HSPCs. (F) Representative H2B-GFP histograms of ES HSCs isolated from 5-FU–perturbed and control mice (supplemental Figure 3C-E). (G-M) Fgd5ZsGreen:CreERT2/R26LSL-tdRFP mice were TAM-induced and perturbed with either 2 Gy γ-radiation (n = 4), 5-FU (n = 5) or left untreated (n = 5). Experiment scheme (G), PB lymphocyte count 7 days after irradiation (H), PB reticulocyte numbers 7 days after 5-FU (I), and ratios of relative BM compartment sizes (% cells among total lin− BM cells) between perturbed and control (dotted line) animals (J-K) are shown. (L) Percentages of RFP+ ES HSCs. (M) Fraction of RFP-labeled cells relative to ES HSCs (significant differences between untreated and either 5-FU–perturbed or irradiated mice was calculated, all comparisons between unperturbed and irradiated mice are not significant). (N-O) R26rtTA/Col1A1H2B-GFP animals were doxycycline (DOX)–pulsed and exposed to either 2 Gy (n = 5), 5-FU (n = 5), or left untreated (n = 6). Experiment scheme (N) and numbers of additional divisions in response to perturbation (O) are shown (supplemental Figure 3C-E). (P) Transformation of data shown in panel M to visualize the net effects of myeloablation on label propagation. Mean (dark gray line) and variance (standard deviation [SD], shaded area) of control mice (n = 41 from 8 independent experiments) are shown (supplemental Figure 3F-H). CMP, common myeloid progenitor; Ery, erythrocyte; MkP, megakaryocyte progenitor; Neut, neutrophil; Plt, platelet; PMN, polymorphonuclear cell.

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