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Blood, Vol. 113, Issue 12, 2805-2815, March 19, 2009
Selective roles for antiapoptotic MCL-1 during granulocyte development and macrophage effector function
Blood Steimer et al.
113: 2805
Supplemental materials for: Steimer et al
Files in this Data Supplement:
- Figure S1. Increased infections in myeloid-specific Mcl-1–deleted mice (JPG, 91.5 KB)
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(A) Hematoxylin and eosin (H&E) stained sections from Mcl-1f/null plus LysM-Cre animal with observable infection. Infections were typically observed on the head and typically involved the ear as depicted in this 2× image. (B) Depicts a 40× enlarged region (boxed in A) demonstrating the extensive cellular infiltrate consisted of lymphocytes, plasma cells, macrophages and few eosinophils. Neutrophils were notably absent from all lesions. (C) Analysis of Mcl-1 genomic deletion by LysM-Cre. Genomic DNA purified from FACS sorted BM neutrophils cells from Mcl-1f/wt plus LysM-Cre or sorted myeloid progenitors from Mcl-1f/null plus LysM-Cre were subjected to polymerase chain reaction to amplify the flox allele (LoxP specific primers), null allele (βGalactosidase-Neomycin fusion gene specific primers) and the deleted allele. CMP, GMP, and MEP were purified from c-Kit+ Lineage-negative BM cells on basis of FcγRII/III and CD34 expression.
- Figure S2. Extramedullary hematopoiesis in myeloid-specific Mcl-1–deleted mice (JPG, 209 KB)
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(A) H&E stained spleen sections from littermate control (Mcl-1f/wt) and Mcl-1-deleted mice (Mcl-1f/null plus LysM-Cre). Mcl-1-deleted mice exhibit extensive extramedullary hematopoiesis and disorganized splenic architecture. (Images taken with 40× objective). (B) Flow cytometric analysis of splenocytes from littermate control and Mcl-1-deleted mice stained for Mac-1 and Gr1. Upper right quadrant represents mature neutrophils and the lower right quadrant immature myeloid precursors.
- Figure S3. Bim-conditional mouse model schematic and genomic analyses (JPG, 76 KB)
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(A) Schematic diagram of the targeting of the Bim genomic locus. Black bars denote Bim exons, H denotes HindIII restriction enzyme sites, Neo refers to the Neomycin resistance gene expressed under control of the mouse PGK promoter. (B) Southern blotting of targeted Bim embryonic stem (ES) cells and F1 progeny mice. Blots are digested with HindIII and probed with the Bim 5′ external homology probe. The fragments are as follows: 11kb is the Bimflox-Neo allele, 9.4 kb is the Bimflox allele, 5.4 kb is the Bim locus after Cre-deletion, and the 2.4 kb is the Bim wild-type allele.
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