The GATA2-deficient BMMCs show characteristics of immature myeloid-like cells. (A) (Upper) BMMCs prepared from ΔCF/+ and ΔCF/ΔCF mice were stained with CD11b and Ly6G/C antibodies and analyzed by FACS after 10 days of 4-OHT treatment. The CD11b⁺/Ly6G/C⁻ and CD11b⁺/Ly6G/C⁺ cells were gated as RI and RII fractions, respectively. (Lower) Average percentages and SD of the RI (left) fractions in the ΔCF/+ (0.2 ± 0.2%) and ΔCF/ΔCF BMMCs (40.7 ± 5.8%) and RII (right) fractions in the ΔCF/+ (0.3 ± 0.1%) and ΔCF/ΔCF(19.7 % ± 1.5%) BMMCs are shown. N = 4. (B) (Left) Representative cytospin preparations of the RI (upper) and RII (lower) cells stained with Wright-Giemsa. Original magnification, ×400. (Right) Mean values of N/C ratio of the RI and RII cells. N = 30. (C) Representative flow cytometric plots of neutrophils (MHCclassII⁻Ly6G⁺Ly6G/C⁺), monocytes (CD11b^highF4/80⁺), basophils (c-Kit⁻FcεRIα⁺CD11b^intCD49b⁺), and eosinophils (CD34⁻CD11b^intCCR3⁺Siglec-F⁺) from ΔCF/ΔCF BMMCs after 0, 5, and 10 days of 4-OHT treatment. (D) The results of the Q-PCR analyses of the indicated genes in ΔCF/ΔCF BMMCs after 0, 5, and 10 days of 4-OHT treatment. N = 4. (E) The expression of the GATA2, GATA1, MITF, CD11b, MPO, C/EBPα, c-Kit, and Lamin-B (control) proteins was analyzed by western blot analysis. Whole cell lysates were isolated from ΔCF/ΔCF BMMCs treated with 4-OHT for 0, 5, and 10 days. The arrowheads indicate the positions of the wild-type and ΔCF GATA2 protein and the p42 and p30 isoforms of C/EBPα. (F) The results of the Q-PCR analyses of basophil-enriched genes in the ΔCF/ΔCF BMMCs after 0, 5, and 10 days of 4-OHT treatment. N = 4. For B, D, and F. *P < .05, **P < .01, ns, not significant. For C-F, samples prepared after 0, 5, and 10 days of 4-OHT treatment are indicated by D0, D5, and D10, respectively.