Figure 4.

Reduced CFU-GM formation and proliferation of c-Kit⁺ cells induced by SCF in Fpr2^−/− mice. A and B, CFU assays. BM cells from WT and Fpr2^−/− mice were treated with the EasySep® mouse hematopoietic progenitor cell enrichment mixture. The enriched cells were cultured in MethoCult® M3434 medium. BFU-E, CFU-GM, and CFU-GEMM were scored, and representative photographs were taken on day 14. A, representative photographs for CFU-GEMM, BFU-E, and CFU-GM from WT and Fpr2^−/− mouse BM. Scale bars = 30 μm. B, cumulative results for CFU-GEMM, BFU-E, and CFU-GM from WT and Fpr2^−/− mice. C–E, BM cells were harvested, and c-Kit⁺ cells were isolated by incubation with anti-mouse c-Kit-FITC Ab, followed by anti-FITC MicroBeads (Miltenyi Biotec). c-Kit⁺ cells were then triple-seeded in 96-well plates in 100 μl of RPMI 1640 (5 × 10⁴/ml) in the presence or absence of SCF (10 ng/ml). alamarBlue was added (10 μl/well) for measurement at 570 nm and 600 nm at the indicated time points. C, c-Kit⁺ cell proliferation stimulated by SCF. *, p < 0.05. D, proliferation of c-Kit⁺ cells from WT mice in the presence or absence of SCF (10 ng/ml). *, p < 0.05. E, the proliferation of c-Kit⁺ cells from Fpr2^−/− mice in the presence or absence of SCF (10 ng/ml). *, p < 0.05. c-Kit⁺ cells isolated from 10 mice/group were pooled with three independent experiments.