Figure 2
Figure 2. Induction of TRF1 deletion depletes HSC and progenitor cells and leads to increased compensatory proliferation. (A) Representative FACS analysis of the HSC and progenitor cell populations. Single cells in panel Ai were further gated based on forward and side scatter (Aii). Including only viable cells without DAPI incorporation, BMMCs of (Aii) were separated on the basis of being lineage negative and IL-7 receptor positive or negative (Aiii). IL-7 receptor–positive cells were further analyzed based on the sca-1 and c-kit staining (Avi). Common lymphoid progenitors (CLP) were identified as sca-1 and c-kit low(+) cells. Lin and IL-7 receptor–negative cells were further distinguished on the basis of sca-1 and c-kit staining (Aiv). HSCs were identified as c-kit(+), sca-1(+). C-kit(+), sca-1(−) progenitor cells were further differentiated in (Av) on the basis of CD34 and Fc-receptor staining. CD34 and Fc-receptor low(+) cells were identified as megakaryocyte-erythrocyte progenitor cells (MEP), CD34(+), Fc-receptor low–positive cells as common myeloid progenitors (CMP),and CD34(+),Fc-receptor(+) cells were identified as granulocyte-macrophage progenitors (GMP). (B) Quantification of the percentage of the hematopoietic stem and progenitor cell subpopulations. The respective percentage was calculated in relation to the number of cells gated for forward and side scatter in panel Aii. Mice without pI-pC–induced Cre expression are represented by dark gray bars and mice undergoing pI-C injections by light gray bars. Two-sided t test was used for statistical comparison between TRF1flox/floxMx1-wt and TRF1flox/floxMx1-Cre group, student paired t test for statistical comparison within the respective group (untreated vs treated). (C) Representative FACS analysis of BrdU incorporation after 2-hour pulse labeling in TRF1flox/floxMx1-wt and TRF1flox/floxMx1-Cre mice undergoing Cre induction. BMMCs were gated for singlet cells and forward and side scatter (FACS scatter gram not shown) and then further separated on the basis of BrdU and propidium iodide staining. (D) Quantification of BrdU-positive cells. The respective percentage was calculated in relation to the number of cells gated for forward and side scatter. Mice without pI-pC–induced Cre expression are represented by dark gray bars and mice undergoing pI-C injections by light gray bars. Two-sided t test and student paired t test was used for statistical comparison between respective TRF1flox/floxMx1-wt and TRF1flox/floxMx1-Cre subgroups.

Induction of TRF1 deletion depletes HSC and progenitor cells and leads to increased compensatory proliferation. (A) Representative FACS analysis of the HSC and progenitor cell populations. Single cells in panel Ai were further gated based on forward and side scatter (Aii). Including only viable cells without DAPI incorporation, BMMCs of (Aii) were separated on the basis of being lineage negative and IL-7 receptor positive or negative (Aiii). IL-7 receptor–positive cells were further analyzed based on the sca-1 and c-kit staining (Avi). Common lymphoid progenitors (CLP) were identified as sca-1 and c-kit low(+) cells. Lin and IL-7 receptor–negative cells were further distinguished on the basis of sca-1 and c-kit staining (Aiv). HSCs were identified as c-kit(+), sca-1(+). C-kit(+), sca-1(−) progenitor cells were further differentiated in (Av) on the basis of CD34 and Fc-receptor staining. CD34 and Fc-receptor low(+) cells were identified as megakaryocyte-erythrocyte progenitor cells (MEP), CD34(+), Fc-receptor low–positive cells as common myeloid progenitors (CMP),and CD34(+),Fc-receptor(+) cells were identified as granulocyte-macrophage progenitors (GMP). (B) Quantification of the percentage of the hematopoietic stem and progenitor cell subpopulations. The respective percentage was calculated in relation to the number of cells gated for forward and side scatter in panel Aii. Mice without pI-pC–induced Cre expression are represented by dark gray bars and mice undergoing pI-C injections by light gray bars. Two-sided t test was used for statistical comparison between TRF1flox/floxMx1-wt and TRF1flox/floxMx1-Cre group, student paired t test for statistical comparison within the respective group (untreated vs treated). (C) Representative FACS analysis of BrdU incorporation after 2-hour pulse labeling in TRF1flox/floxMx1-wt and TRF1flox/floxMx1-Cre mice undergoing Cre induction. BMMCs were gated for singlet cells and forward and side scatter (FACS scatter gram not shown) and then further separated on the basis of BrdU and propidium iodide staining. (D) Quantification of BrdU-positive cells. The respective percentage was calculated in relation to the number of cells gated for forward and side scatter. Mice without pI-pC–induced Cre expression are represented by dark gray bars and mice undergoing pI-C injections by light gray bars. Two-sided t test and student paired t test was used for statistical comparison between respective TRF1flox/floxMx1-wt and TRF1flox/floxMx1-Cre subgroups.

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