Figure 4.
CD11c deficiency affects repopulation of HSPCs. (A-F) Mixed chimeric mouse study. Chimeric mice were made by transplanting the mixture of CD45.1 WT and CD45.2 WT BM cells or CD45.1 WT and CD45.2 CD11c−/− BM cells at the ratio of 1:1 into lethally irradiated recipient mice. (A) At 10 weeks post-BM transplantation, peripheral blood from recipient mice was collected and subjected to flow cytometry analysis. Shown are representatives of 5 mice in both groups. (B-C) At 12 weeks, BM cells from recipient mice were collected and subjected to flow cytometry analysis. (B) Left panel, Representative flow cytometry plots of lineage-negative cells, LSKs, and CMPs from different originations differentiated by congenic markers. Right panel, Cumulative absolute number of LSKs at various time points post-BM transplantation. (C) Representative flow cytometry plots of BM neutrophils from different originations differentiated by congenic markers at 12 weeks post-BM transplantation. Shown are representatives of 5 mice in both groups. (D-E) Proliferation measured by in vivo BrdU labeling. At 2 weeks post-BM transplantation, BrdU pulse injection was performed to examine the proliferation. (D) Representative flow cytometry plots. (E) The frequency of BrdU+ in LSKs in WT/WT chimera or in WT/ CD11c−/− chimera. Line indicates an individual chimeric mouse. (F) Z-VAD-fmk treatment of WT/ CD11c−/− chimera. Starting at 1 week after BM transplantation, chimeric mice were treated with either vehicle (5% DMSO in saline, IP) or Z-VAD (10 mg/kg, dissolved in 5% DMSO in saline, IP) daily for 4 days. Then, the mice were euthanized and BM LSKs and CMPs were counted and analyzed by flow cytometry. (Left 2 panels) Cell number. (Right panel) Ratio of CD11c−/− derived CD45.2 positive cells in LSK or CMP. (A-C) Representatives of 3 to 5 independent experiments with the same pattern. (D-F) Representatives of 2 independent experiments with the same pattern. Symbols indicate individual mice. Statistical significance was examined by the Student t test (E-F). *P < .05.

CD11c deficiency affects repopulation of HSPCs. (A-F) Mixed chimeric mouse study. Chimeric mice were made by transplanting the mixture of CD45.1 WT and CD45.2 WT BM cells or CD45.1 WT and CD45.2 CD11c−/− BM cells at the ratio of 1:1 into lethally irradiated recipient mice. (A) At 10 weeks post-BM transplantation, peripheral blood from recipient mice was collected and subjected to flow cytometry analysis. Shown are representatives of 5 mice in both groups. (B-C) At 12 weeks, BM cells from recipient mice were collected and subjected to flow cytometry analysis. (B) Left panel, Representative flow cytometry plots of lineage-negative cells, LSKs, and CMPs from different originations differentiated by congenic markers. Right panel, Cumulative absolute number of LSKs at various time points post-BM transplantation. (C) Representative flow cytometry plots of BM neutrophils from different originations differentiated by congenic markers at 12 weeks post-BM transplantation. Shown are representatives of 5 mice in both groups. (D-E) Proliferation measured by in vivo BrdU labeling. At 2 weeks post-BM transplantation, BrdU pulse injection was performed to examine the proliferation. (D) Representative flow cytometry plots. (E) The frequency of BrdU+ in LSKs in WT/WT chimera or in WT/ CD11c−/− chimera. Line indicates an individual chimeric mouse. (F) Z-VAD-fmk treatment of WT/ CD11c−/− chimera. Starting at 1 week after BM transplantation, chimeric mice were treated with either vehicle (5% DMSO in saline, IP) or Z-VAD (10 mg/kg, dissolved in 5% DMSO in saline, IP) daily for 4 days. Then, the mice were euthanized and BM LSKs and CMPs were counted and analyzed by flow cytometry. (Left 2 panels) Cell number. (Right panel) Ratio of CD11c−/− derived CD45.2 positive cells in LSK or CMP. (A-C) Representatives of 3 to 5 independent experiments with the same pattern. (D-F) Representatives of 2 independent experiments with the same pattern. Symbols indicate individual mice. Statistical significance was examined by the Student t test (E-F). *P < .05.

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