Figure 2
Figure 2. Enhanced myelopoiesis in Icsbp−/−Nf1+/− mice. (A) Highest total leukocyte counts from peripheral blood in Icsbp−/−Nf1+/− mice. Total leukocyte counts were determined using an automated hematologic cell counter from the peripheral blood (tail vein) from mice 2 to 4 months (young, n = 26) and 7 to 9 months of age (old, n = 16). Dots represent individual measurements; lines, medians; the level of significance comparing Icsbp−/−Nf1+/− and Icsbp−/−Nf1+/+ mice is shown above dots. Nonparametric test was used (Mann-Whitney U test) to calculate level of significance. (B) Highest neutrophil granulocyte counts in peripheral blood in Icsbp−/−Nf1+/− mice. Blood smears (n = 5) were prepared from peripheral blood, and total numbers of neutrophil granulocytes were determined blindly after May-Gruenwald-Giemsa staining. FACS analysis was performed after lysis of erythrocytes. Cells staining high with Gr-1 and CD11b were identified as neutrophil granulocytes. Absolute numbers of neutrophil granulocytes were calculated multiplying numbers of total leukocytes with the percentage of cells staining for Gr-1 and CD11b (n = 5). Dots represent individual measurements; lines, medians; the level of significance (Mann-Whitney U test) comparing Icsbp−/−Nf1+/− and Icsbp−/−Nf1+/+ mice is shown above dots. (C) Highest splenomegaly in Icsbp−/−Nf1+/− mice. Spleens and total body weights from mice 2 to 4 months (young, n = 25) and 7 to 9 months (old, n = 12) were determined using a precision balance. Dots represent individual measurements; lines, medians; the level of significance (Mann-Whitney U test) comparing Icsbp−/−Nf1+/− and Icsbp−/−Nf1+/+ mice is shown above dots. (D) Infiltration of neutrophil granulocytes into lymph nodes. Percentages of neutrophil granulocytes were determined using flow cytometry staining in BM, spleen, and lymph nodes from mice 4 months of age. Cells were pregated in a live gate and resolved for staining of Gr-1 and CD11b. Numbers above squares represent mean ± SD of n = 5 individual mice. Statistical significance (Mann-Whitney U test): *between Icsbp−/−Nf1+/+ and Icsbp+/+Nf1+/+; **between Icsbp−/−Nf1+/− and Icsbp+/+Nf1+/−.

Enhanced myelopoiesis in Icsbp−/−Nf1+/− mice. (A) Highest total leukocyte counts from peripheral blood in Icsbp−/−Nf1+/− mice. Total leukocyte counts were determined using an automated hematologic cell counter from the peripheral blood (tail vein) from mice 2 to 4 months (young, n = 26) and 7 to 9 months of age (old, n = 16). Dots represent individual measurements; lines, medians; the level of significance comparing Icsbp−/−Nf1+/− and Icsbp−/−Nf1+/+ mice is shown above dots. Nonparametric test was used (Mann-Whitney U test) to calculate level of significance. (B) Highest neutrophil granulocyte counts in peripheral blood in Icsbp−/−Nf1+/− mice. Blood smears (n = 5) were prepared from peripheral blood, and total numbers of neutrophil granulocytes were determined blindly after May-Gruenwald-Giemsa staining. FACS analysis was performed after lysis of erythrocytes. Cells staining high with Gr-1 and CD11b were identified as neutrophil granulocytes. Absolute numbers of neutrophil granulocytes were calculated multiplying numbers of total leukocytes with the percentage of cells staining for Gr-1 and CD11b (n = 5). Dots represent individual measurements; lines, medians; the level of significance (Mann-Whitney U test) comparing Icsbp−/−Nf1+/− and Icsbp−/−Nf1+/+ mice is shown above dots. (C) Highest splenomegaly in Icsbp−/−Nf1+/− mice. Spleens and total body weights from mice 2 to 4 months (young, n = 25) and 7 to 9 months (old, n = 12) were determined using a precision balance. Dots represent individual measurements; lines, medians; the level of significance (Mann-Whitney U test) comparing Icsbp−/−Nf1+/− and Icsbp−/−Nf1+/+ mice is shown above dots. (D) Infiltration of neutrophil granulocytes into lymph nodes. Percentages of neutrophil granulocytes were determined using flow cytometry staining in BM, spleen, and lymph nodes from mice 4 months of age. Cells were pregated in a live gate and resolved for staining of Gr-1 and CD11b. Numbers above squares represent mean ± SD of n = 5 individual mice. Statistical significance (Mann-Whitney U test): *between Icsbp−/−Nf1+/+ and Icsbp+/+Nf1+/+; **between Icsbp−/−Nf1+/− and Icsbp+/+Nf1+/−.

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