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Blood, Vol. 114, Issue 10, 2107-2120, September 3, 2009
Gata1 expression driven by the alternative HS2 enhancer in the spleen rescues the hematopoietic failure induced by the hypomorphic Gata1low mutation
Blood Migliaccio et al.
114: 2107
Supplemental materials for: Migliaccio et al
Files in this Data Supplement:
- Table S1. Number of splenectomized mine analyzed at each time point (PDF, 14 KB)
- Table S2. Gene expression and analysis of single colonies derived from the bone marrow of untreated wild type and Gata1low∕0 males or from untreated and splenectomized Gata1low∕+ females (PDF, 47.5 KB)
- Table S3. Total number of cKitpos cells present in the bone marrow and spleen of wild type and Gata1low littermates (PDF, 14.2 KB)
- Table S4. Blood values in NOD/SCID mice either untreated or after 1 months of being injected with 1 × 106 bone marrow cells from Gata1low mice (PDF, 13.4 KB)
- Figure S1. Design of the splenectomy experiments (JPG, 138 KB)
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Hemizygous Gata1low∕0 males and heterozygous Gata1low∕+ females were splenectomized at 6 month of age, i.e., in the Pre-MF phase, and were sacrificed 3, 6 and 9–10 months later for evidence of disease progression12,14. End points of these experiments were represented by blood values (Hct, Plt and WBC counts, presence of tear drop poikylocytes), bone marrow parameters (marrow cellularity, MK frequency, levels of Gata1 expression in MK by immunohistochemistry, marrow fibrosis (MF), osteogenesis and frequency of progenitor cells expressing the wild-type or the Gata1low allele). The number and survival rate of the splenectomized mice are summarized Table S1.
- Figure S2. Splenectomy increases the cellularity of the marrow and induces elevation in blood platelet counts in Gatalow∕+ (JPG, 248 KB)
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A and B indicate cellularity and frequency of MK in the marrow while C and D indicate platelet counts and hematocrit (Hct) of Gatalow∕+ mice at different time points following splenectomy. The total cell number in the marrow was evaluated by counting the number of nucleated cells flushed from the femoral cavity. In addition, the frequency of MK in the different specimens was quantified by counting the number of MK per mm2 of marrow sections stained with hematoxylin-eosin (Fig. 2A). The corresponding age of the mice is reported on the top, for comparison. Results are expressed as a mean (±SD) of at least 5–7 animals per experimental group and are compared to those expressed by untreated Gata1low (grey area) littermates of comparable age, as previously published.14 Values statistically significant (p< 0.05 or .01) as compared to untreated controls are indicated by * or **. Results obtained with 6–7 month old wild-type mice are presented as columns, for comparison.
- Figure S3. Flow cytometrical analyses for CD41/CD61 expression (on the right) and for the corresponding isotype controls (on the left) of cells from the blood, bone marrow, spleen and liver of a representative NOD/SCID mouse transplanted with marrow cells from Gata1low mice (JPG, 225 KB)
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