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. 2017 Aug 11;24(12):2032–2043. doi: 10.1038/cdd.2017.125

Figure 1.

Figure 1

Reduction in MCL-1 levels causes a significant albeit minor loss of certain blood cell subsets. (a) Representative example of intracellular FACS staining for MCL-1, BIM, BCL-2 and BCL-XL in B220+ gated B cells from the blood of wild-type (wt) and Mcl-1+/− mice (left panel). Data in the right panel represent relative mean fluorescence intensity (MFI) ±S.E.M. in wild-type (n≥5) and Mcl-1+/− mice (n≥5). ***P<0.001 (Student's t-test, two tailed, unpaired). (b) Total white blood cell (WBC), lymphocyte (LC), RBC numbers, haemoglobin content (HGB) and haematocrit (HCT) in peripheral blood of wild-type (n=42) and Mcl-1+/− (n=54) mice. Data represent mean ±S.E.M. *P<0.05, **P<0.01, ***P<0.001 (Student's t-test, two tailed, unpaired). (c) Representative examples of flow cytometric analysis of T cells, B cells (left panel) and myeloid cells (right panel) identified by surface staining for TCRβ/B220 and MAC-1/GR-1, respectively, in the peripheral blood of wild-type and Mcl-1+/− mice. (d) Data are presented as mean ±S.E.M. of total numbers of the indicated cell types in peripheral blood, spleen and bone marrow (BM) of wild-type (n=7) and Mcl-1+/− (n=7) mice. P>0.5 (n.s.) (Student's t-test, two tailed, unpaired, comparing wild-type with Mcl-1+/− mice). (e) Histological analysis of H&E-stained sections of the BM (sternum) of wild-type and Mcl-1+/− mice. (f) Representative examples of flow cytometric analysis of the indicated B-cell populations identified by staining for IgM/B220. I=pro-B/pre-B, II=immature B; III=transitional B; IV= mature B cells (left panel). Data are presented as mean ±S.E.M. of the total numbers of the indicated cell subsets in the bone marrow (total cell count per one femur) of wild-type (n=7) and Mcl-1+/− (n=7) mice (right panel). P>0.5 (n.s.) (Student's t-test, two tailed, unpaired, comparing wild-type with Mcl-1+/− mice). (g) Representative examples of flow cytometric analysis of thymic T-cell populations identified by staining for CD4/CD8 (left panel). Data are presented as mean ±S.E.M. of total numbers of the indicated cell subsets in the thymi from wild-type (n=7) and Mcl-1+/− (n=7) mice (right panel). P>0.5 (n.s.) (Student's t-test, two tailed, unpaired, comparing wild-type with Mcl-1+/− mice). DN=double negative; DP=double positive; SP single positive. (h) Representative examples of flow cytometric analysis of long-term haematopoietic stem cells (LT-HSC) identified by staining with lineage marker cocktail antibodies, and antibodies against c-KIT, SCA-1, CD48 and CD150. Lin- (lineage negative), LSK (Lin−c-KIT+SCA-1+ (left panel)). Data are presented as mean ±S.E.M. of the total numbers of the indicated cell subsets in the bone marrow (total cell count per one femur) of wild-type (n=5) and Mcl-1+/− (n=5) mice (right panel). P>0.5 (n.s.) (Student's t-test, two tailed, unpaired, comparing wild-type with Mcl-1+/− mice)