Figure 2
Figure 2. Basophils as a major producer of CCL3 in the BM. (A) Femoral and tibial BM-derived HPCs were injected into the right tibial BM cavity of 2 Gy irradiated CD45.1+ congenic mice. The c-kit and CCL3 expression on the CD45.2+MPO+lineagelow cells (R1 and R2) was determined. Percentages of CD45.2+MPO+ donor-derived myeloid cells and c-kitlowCCL3+ cells are shown in the upper and lower panels, respectively. Representative results from 3 independent experiments are shown. (B) LineagelowCD16/32highc-kitlow cells and lineagelowCD16/32highc-kithigh granulocyte macrophage progenitor were isolated from femoral and tibial BMs using a FACSAria Cell Sorter. The obtained cells were stained with Wright and Giemsa staining solution, and representative results from 3 independent experiments are shown with a scale bar of 10 μm. Images were obtained using a BX50 microscope (Olympus) using a ×40 objective lens. CCL3 expression was determined using quantitative real-time polymerase chain reaction analysis. Means ± SD from 3 independent experiments are given. *P < .05; Tukey-Kramer test. (C) CCL3 expression in c-kitlowCD49b+FcεR1+ cells (R1 and R2) in the tibial BM of untreated or BM chimeric mice at 8 weeks after transplantation. Rat IgG2a was used as an isotype control. Representative results from 4 independent experiments are shown. (D) CCL3 expression in CD200R3+FcεR1+CD34+FSChigh (BaP1), CD34+FSClow (BaP2), and CD34lowFSClow cells (Ba) in the tibial BM and PB. Rat IgG2a was used as an isotype control. Representative results from 3 independent experiments are shown. The mean fluorescence intensity of CCL3 in each population was compared with that in PB Ba. **P < .01; N.S., no significant difference by Dunnett’s test.

Basophils as a major producer of CCL3 in the BM. (A) Femoral and tibial BM-derived HPCs were injected into the right tibial BM cavity of 2 Gy irradiated CD45.1+ congenic mice. The c-kit and CCL3 expression on the CD45.2+MPO+lineagelow cells (R1 and R2) was determined. Percentages of CD45.2+MPO+ donor-derived myeloid cells and c-kitlowCCL3+ cells are shown in the upper and lower panels, respectively. Representative results from 3 independent experiments are shown. (B) LineagelowCD16/32highc-kitlow cells and lineagelowCD16/32highc-kithigh granulocyte macrophage progenitor were isolated from femoral and tibial BMs using a FACSAria Cell Sorter. The obtained cells were stained with Wright and Giemsa staining solution, and representative results from 3 independent experiments are shown with a scale bar of 10 μm. Images were obtained using a BX50 microscope (Olympus) using a ×40 objective lens. CCL3 expression was determined using quantitative real-time polymerase chain reaction analysis. Means ± SD from 3 independent experiments are given. *P < .05; Tukey-Kramer test. (C) CCL3 expression in c-kitlowCD49b+FcεR1+ cells (R1 and R2) in the tibial BM of untreated or BM chimeric mice at 8 weeks after transplantation. Rat IgG2a was used as an isotype control. Representative results from 4 independent experiments are shown. (D) CCL3 expression in CD200R3+FcεR1+CD34+FSChigh (BaP1), CD34+FSClow (BaP2), and CD34lowFSClow cells (Ba) in the tibial BM and PB. Rat IgG2a was used as an isotype control. Representative results from 3 independent experiments are shown. The mean fluorescence intensity of CCL3 in each population was compared with that in PB Ba. **P < .01; N.S., no significant difference by Dunnett’s test.

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