Figure 2
Figure 2. Deletion of miR-142 results in abnormal development of lymphoid cells. (A-B) Dramatic decrease in the number of B1 B cells in miR-142−/− mice. (A) FACS analysis of lymphocytes in the peritoneal cavity (PC) of WT and KO mice with anti-B220 and anti-CD19 specific antibodies. B1 (CD19+B220lo) and B2 (CD19+B220hi) cell populations are gated, and numbers indicate the percentage of cells in the gate. (B) Total B1 B-cell numbers in the PC of WT (n = 5) and KO (n = 6) mice. (C-D) Increased numbers of MZ-like B cells in miR-142−/− spleens. (C) FACS analysis of CD19+ B cells in WT and KO spleens with anti-CD21 and anti-CD23 antibodies. FO (CD21intCD23+), MZ (CD21hiCD23lo), and newly formed (NFB) (CD21intCD23lo) B-cell subsets are gated, and numbers indicate the percentage of cells in the gate. (D) Total numbers of FO, MZ, and NFB B cells in WT (n = 3) and KO (n = 3) spleens. (E-G) Altered B-cell maturation in miR-142−/− mice. (E) FACS analysis of WT and KO splenocytes with anti-B220 and anti-CD93 antibodies. Cells were pregated on lymphocytes. Immature (IM) (B220+CD93+) and mature (M) (B220+CD93−) B-cell populations are gated, and the numbers indicate the percentage of cells in the gate. (F) FACS analysis of B220+CD93+ immature B cells from WT and KO spleens with anti-IgM and anti-CD23 antibodies. Transitional T1 (IgM+CD23−), T2 (IgM+CD23+), T3 (IgM-CD23+), and precursor (pre-B) (IgM−CD23−) B cells are gated, and numbers indicate the percentage of cells in the gate. (G) Total numbers of pre-B, immature, transitional (T1-T3), and mature B cells in WT (n = 4) and KO (n = 4) spleens. (H-J) Decrease in peripheral T-cell numbers, but normal central T cell development in miR-142−/− mice. (H) FACS analysis of splenocytes in WT and KO mice with anti-CD3ε antibodies. T cells (CD3+) are gated, and numbers indicate the percentage of cells in the gate. (I) Total number of T cells in WT (n = 8) and KO (n = 8) spleens. (J) FACS analysis of WT and KO thymocytes with anti-CD4 and anti-CD8 antibodies. Numbers indicate the percentage of cells in the quadrants. (K-M) Elevated B-/T-cell ratio in miR-142−/− mice. (K) FACS analysis of WT and KO splenocytes with anti-CD3ε and anti-CD19 antibodies. B cells (CD19+) and T cells (CD3+) are gated, and numbers indicate the percentage of cells in the gate. (L) The ratio of B to T cells in WT (n = 3) and KO (n = 3) spleens. (M) Immunohistochemical analysis of spleen sections from WT and miR-142 KO mice with anti-B220 (red stain) and anti-CD3 (brown stain) antibodies. Scale bar, 100 μm. Note abnormal spleen architecture in the KO: smaller T-cell zone and less organized, less compact B-cell zone. Results are shown as means ± SD and are representative of at least 3 independent experiments. P values were calculated using Student t test. *P ≤ .05; **P ≤ .01; ***P ≤ .001; ****P ≤ .0001. B, B-cell follicle; ns, not significant; T, T-cell zone.

Deletion of miR-142 results in abnormal development of lymphoid cells. (A-B) Dramatic decrease in the number of B1 B cells in miR-142−/− mice. (A) FACS analysis of lymphocytes in the peritoneal cavity (PC) of WT and KO mice with anti-B220 and anti-CD19 specific antibodies. B1 (CD19+B220lo) and B2 (CD19+B220hi) cell populations are gated, and numbers indicate the percentage of cells in the gate. (B) Total B1 B-cell numbers in the PC of WT (n = 5) and KO (n = 6) mice. (C-D) Increased numbers of MZ-like B cells in miR-142−/− spleens. (C) FACS analysis of CD19+ B cells in WT and KO spleens with anti-CD21 and anti-CD23 antibodies. FO (CD21intCD23+), MZ (CD21hiCD23lo), and newly formed (NFB) (CD21intCD23lo) B-cell subsets are gated, and numbers indicate the percentage of cells in the gate. (D) Total numbers of FO, MZ, and NFB B cells in WT (n = 3) and KO (n = 3) spleens. (E-G) Altered B-cell maturation in miR-142−/− mice. (E) FACS analysis of WT and KO splenocytes with anti-B220 and anti-CD93 antibodies. Cells were pregated on lymphocytes. Immature (IM) (B220+CD93+) and mature (M) (B220+CD93−) B-cell populations are gated, and the numbers indicate the percentage of cells in the gate. (F) FACS analysis of B220+CD93+ immature B cells from WT and KO spleens with anti-IgM and anti-CD23 antibodies. Transitional T1 (IgM+CD23−), T2 (IgM+CD23+), T3 (IgM-CD23+), and precursor (pre-B) (IgM−CD23−) B cells are gated, and numbers indicate the percentage of cells in the gate. (G) Total numbers of pre-B, immature, transitional (T1-T3), and mature B cells in WT (n = 4) and KO (n = 4) spleens. (H-J) Decrease in peripheral T-cell numbers, but normal central T cell development in miR-142−/− mice. (H) FACS analysis of splenocytes in WT and KO mice with anti-CD3ε antibodies. T cells (CD3+) are gated, and numbers indicate the percentage of cells in the gate. (I) Total number of T cells in WT (n = 8) and KO (n = 8) spleens. (J) FACS analysis of WT and KO thymocytes with anti-CD4 and anti-CD8 antibodies. Numbers indicate the percentage of cells in the quadrants. (K-M) Elevated B-/T-cell ratio in miR-142−/− mice. (K) FACS analysis of WT and KO splenocytes with anti-CD3ε and anti-CD19 antibodies. B cells (CD19+) and T cells (CD3+) are gated, and numbers indicate the percentage of cells in the gate. (L) The ratio of B to T cells in WT (n = 3) and KO (n = 3) spleens. (M) Immunohistochemical analysis of spleen sections from WT and miR-142 KO mice with anti-B220 (red stain) and anti-CD3 (brown stain) antibodies. Scale bar, 100 μm. Note abnormal spleen architecture in the KO: smaller T-cell zone and less organized, less compact B-cell zone. Results are shown as means ± SD and are representative of at least 3 independent experiments. P values were calculated using Student t test. *P ≤ .05; **P ≤ .01; ***P ≤ .001; ****P ≤ .0001. B, B-cell follicle; ns, not significant; T, T-cell zone.

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