Figure 6.
Figure 6. Autonomous repopulation of recipient BM Mφs synchronizes with BM engraftment of phenotypic donor HSCs. (A) Representative flow cytometry analysis of naive (no-Tx) and 5 week posttransplant MacGreen recipient BM. LSK cells (black, gate i) consist of CD48+ HPCs (blue, gate ii), CD48negCD150neg MPP (yellow, gate iv), and CD48negCD150+ HSCs (red, gate iii). (B) Enumeration of total number of HPCs (blue bars), MPPs (yellow bars), and HSCs (red bars) in the BM (n = 5-12 mice/time point). Data are presented as mean ± SD. Statistical analysis used a 1-way ANOVA Tukey’s multiple comparison test on donor HSCs (red bars) in transplanted groups, and asterisks represent significant differences in HSC numbers compared with 12 weeks posttransplant. (C) Overlay of results from Figure 1F-G with Figure 6B to align recovery kinetics of posttransplant Mφ numbers (#, left y-axis) with donor HSC numbers (right y-axis) in the BM. Dotted boxed area indicates recipient Mφ population number peak. (D) Enumeration of total number of HPCs (blue bars), MPPs (yellow bars), and HSCs (red bars) in the spleen. Data are presented as mean ± SD. Statistical analysis used a 1-way ANOVA Tukey’s multiple comparison test on donor HPCs (blue bars) in transplanted groups, and asterisks represent significant differences in HPC numbers compared with 2 weeks posttransplant. (E) Overlay of results from Figure 2E with panel D to align recovery kinetics of posttransplant Mφ numbers (#, left y-axis) with donor HPC numbers (right y-axis) in spleen. Dotted boxed area indicates recipient Mφ population number peak. Significance values are designated as *P < .05, **P < .01, and ***P < .001. Flow cytometry plots and histograms are from a representative animal, and the percentage of positive cells is based on the proceeding parent populations. Data are representative of 5 to 13 individual animal samples per time point from either 1 (week 3 and 32), 2 (week 5), or 3 (all other time points) biological replicate experiments. 1 × 106 events were collected from each experiment sample by flow cytometry.

Autonomous repopulation of recipient BM Mφs synchronizes with BM engraftment of phenotypic donor HSCs. (A) Representative flow cytometry analysis of naive (no-Tx) and 5 week posttransplant MacGreen recipient BM. LSK cells (black, gate i) consist of CD48+ HPCs (blue, gate ii), CD48negCD150neg MPP (yellow, gate iv), and CD48negCD150+ HSCs (red, gate iii). (B) Enumeration of total number of HPCs (blue bars), MPPs (yellow bars), and HSCs (red bars) in the BM (n = 5-12 mice/time point). Data are presented as mean ± SD. Statistical analysis used a 1-way ANOVA Tukey’s multiple comparison test on donor HSCs (red bars) in transplanted groups, and asterisks represent significant differences in HSC numbers compared with 12 weeks posttransplant. (C) Overlay of results from Figure 1F-G with Figure 6B to align recovery kinetics of posttransplant Mφ numbers (#, left y-axis) with donor HSC numbers (right y-axis) in the BM. Dotted boxed area indicates recipient Mφ population number peak. (D) Enumeration of total number of HPCs (blue bars), MPPs (yellow bars), and HSCs (red bars) in the spleen. Data are presented as mean ± SD. Statistical analysis used a 1-way ANOVA Tukey’s multiple comparison test on donor HPCs (blue bars) in transplanted groups, and asterisks represent significant differences in HPC numbers compared with 2 weeks posttransplant. (E) Overlay of results from Figure 2E with panel D to align recovery kinetics of posttransplant Mφ numbers (#, left y-axis) with donor HPC numbers (right y-axis) in spleen. Dotted boxed area indicates recipient Mφ population number peak. Significance values are designated as *P < .05, **P < .01, and ***P < .001. Flow cytometry plots and histograms are from a representative animal, and the percentage of positive cells is based on the proceeding parent populations. Data are representative of 5 to 13 individual animal samples per time point from either 1 (week 3 and 32), 2 (week 5), or 3 (all other time points) biological replicate experiments. 1 × 106 events were collected from each experiment sample by flow cytometry.

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