Figure 2
Figure 2. Aged Runx3 KO mice shows enhanced myeloproliferation with an expanded HSPC compartment. (A) Complete blood counts performed on 18-month-old Runx3 WT (n = 9) and Runx3 KO mice (n = 21). WBC, Hb, and Plt counts are shown. The statistical significance (P value, Student t test) is shown at the top. (B) May-Grünwald-Giemsa staining of PB cells in aged mice. Representative pictures of cells are shown. (C) Number of cells of each lineage in BM of 18-month-old mice. Lineage markers: myeloid, Mac-1+ and Gr-1+; B-cells, B220+CD19+; T cells, CD3+; erythroid, Ter119+. Mean ± standard deviation (SD) is shown (WT, n = 2; KO, n = 4). Three independent experiments were performed. Asterisks represent significant difference (**P < .01, Student t test). (D) BM cellularity of aged mice. Mean ± SD of the numbers of cells in BM are shown (WT, n = 2; KO, n = 4). Asterisks represent significant difference (**P < .01, Student t test). (E) Spleen of aged Runx3 KO mice. (Left) Graphical representation of spleen weight in the aged mice. Mean ± SD is shown (WT, n = 2; KO, n = 4). Asterisk represents significant difference (*P < .05, Student t test). Three independent experiments were performed. (Right) Representative pictures of spleens in the aged mice. (F,H) Flow cytometric analysis of the HSPC compartment in (F) BM and (H) spleen of 18-month-old mice. Representative FACS plots of 200 000 cells gated on viable Lineage− cells are shown. (G,I) Graphical representations of results presented in F and H are shown in G and I, respectively. Mean ± SD of percentage of c-Kit+Sca-1−Lineage− (myeloid progenitors) and KSL within the BM Lineage− population is shown (WT: n = 2; KO: n = 4). Three independent experiments were performed. (J) Time course PB chimerism analysis of recipient mice after BMT. CD45.1/CD45.2 competitive cells were cotransplanted with Runx3 WT or KO BM cells (CD45.2/CD45.2) into CD45.1/CD45.1 sublethally irradiated (8 Gy) mice in a 1:1 ratio. Percentage contribution of CD45.2/CD45.2 donor cells to total PB, myeloid, B-cell, and T-cell populations at indicated time points after BMT are shown. Mean ± SD is shown (WT, n = 3; KO, n = 6). Five of 8 mice receiving Runx3 WT donor cells and 3 of 9 mice receiving Runx3 KO donor cells that failed to show contribution of CD45.2/CD45.2 cells to PB of recipient mice are excluded. Asterisk represents significant differences (*P < .05, Student t test).

Aged Runx3 KO mice shows enhanced myeloproliferation with an expanded HSPC compartment. (A) Complete blood counts performed on 18-month-old Runx3 WT (n = 9) and Runx3 KO mice (n = 21). WBC, Hb, and Plt counts are shown. The statistical significance (P value, Student t test) is shown at the top. (B) May-Grünwald-Giemsa staining of PB cells in aged mice. Representative pictures of cells are shown. (C) Number of cells of each lineage in BM of 18-month-old mice. Lineage markers: myeloid, Mac-1+ and Gr-1+; B-cells, B220+CD19+; T cells, CD3+; erythroid, Ter119+. Mean ± standard deviation (SD) is shown (WT, n = 2; KO, n = 4). Three independent experiments were performed. Asterisks represent significant difference (**P < .01, Student t test). (D) BM cellularity of aged mice. Mean ± SD of the numbers of cells in BM are shown (WT, n = 2; KO, n = 4). Asterisks represent significant difference (**P < .01, Student t test). (E) Spleen of aged Runx3 KO mice. (Left) Graphical representation of spleen weight in the aged mice. Mean ± SD is shown (WT, n = 2; KO, n = 4). Asterisk represents significant difference (*P < .05, Student t test). Three independent experiments were performed. (Right) Representative pictures of spleens in the aged mice. (F,H) Flow cytometric analysis of the HSPC compartment in (F) BM and (H) spleen of 18-month-old mice. Representative FACS plots of 200 000 cells gated on viable Lineage− cells are shown. (G,I) Graphical representations of results presented in F and H are shown in G and I, respectively. Mean ± SD of percentage of c-Kit+Sca-1−Lineage− (myeloid progenitors) and KSL within the BM Lineage− population is shown (WT: n = 2; KO: n = 4). Three independent experiments were performed. (J) Time course PB chimerism analysis of recipient mice after BMT. CD45.1/CD45.2 competitive cells were cotransplanted with Runx3 WT or KO BM cells (CD45.2/CD45.2) into CD45.1/CD45.1 sublethally irradiated (8 Gy) mice in a 1:1 ratio. Percentage contribution of CD45.2/CD45.2 donor cells to total PB, myeloid, B-cell, and T-cell populations at indicated time points after BMT are shown. Mean ± SD is shown (WT, n = 3; KO, n = 6). Five of 8 mice receiving Runx3 WT donor cells and 3 of 9 mice receiving Runx3 KO donor cells that failed to show contribution of CD45.2/CD45.2 cells to PB of recipient mice are excluded. Asterisk represents significant differences (*P < .05, Student t test).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal