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Blood, Vol. 112, Issue 6, 2500-2507, September 15, 2008
FIP1L1/PDGFR synergizes with SCF to induce systemic mastocytosis in a murine model of chronic eosinophilic leukemia/hypereosinophilic syndrome
Blood Yamada et al.
112: 2500
Supplemental materials for: Yamada et al
Files in this Data Supplement:
- Figure S1. F/P positive CEL mice display bone marrow myelofibrosis (JPG, 108 KB)
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Masson trichromic staining. (A-B) High magnification (original magnification, ×400) of longitudinal tibia sections of mice transplanted with either mock-transduced/WT or F/P-transduced/IL-5Tg HSC/P. Collagen infiltrates appear in blue color. Collagen infiltration in mock-transduced/IL5Tg recipient mice was similar to mock-transduced/WT recipients. Bar = 100 µm.
- Figure S2. Morphology of MC from F/P positive CEL mice is abnormal (JPG, 111 KB)
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(A-B) CAE staining of high magnification (original magnification, ×1000) of representative small intestine sections from CEL/HES and control mice (mock-transduced/WT or mock-transduced;IL-5Tg recipients). Intestinal hyperplasia of MC in CEL/HES mice, which show irregular shape (arrows) compared with the round shape of scarce MC in control mice. Bar = 30 µm. (C-D) Light scattergrams (flow cytometry analysis) of BMMC (c-kit+/FcεRI+) transduced with either F/P or empty vector (mock). Flow cytometry analysis was performed at different times of culture with SCF (C) or IL-3 (D). SCF-cultured F/P-expressing BMMC show a much higher property of light side scatter than mock-transduced BMMC cultured with SCF, mock-transduced BMMC cultured with IL-3 or F/P-transduced BMMC cultured with IL-3. Representative example of two or three independent experiments.
- Figure S3. IL-5 overexpression increases the level of intestinal MC and serum MMCP-1 (JPG, 51.4 KB)
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(A) Levels of mast cells in the small intestine. Levels of intestinal mast cells in mice transplanted with mock-transduced/WT, mock-transduced/IL-5Tg, F/P-transduced/WT and F/P-transduced/IL-5Tg HSC/P are presented. MC content was assessed after development of disease (at 4-5 weeks post transplantation) by morphometric analysis of chloroacetate esterase (CAE) stained cells. Results are shown as mean ± SEM from 4-6 mice per group of one representative experiment (n = 3 experiments. *P< 0.01). (B) Levels of cutaneous mast cells in the same mice as in A. Results are shown as mean ± SEM from 3-4 mice per group of one representative experiment, * P< 0.05. C) Levels of serum MMCP-1 in the same groups of mice as in A and B. mMCP-1 serum levels were assessed after development of disease (at 4-5 weeks post transplantation) by ELISA. Each dot represents results from one mouse and the horizontal bars show mean values (n=6-8, *p< 0.001). The results are pooled from two independent experiments.
- Figure S4. Time course of SCF-induced activation of Akt (p-Akt) (JPG, 43.2 KB)
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Results are normalized by the β-actin expression and presented as fold-increase. Results denote the average and SEM of 4 independent experiments, normalized over the 5 min activation of WT MC.
- Figure S5. Time course of SCF-induced activation of Akt, Erk and STAT-5 (JPG, 66.1 KB)
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SCF-induced activation of Akt (p-Akt), Erk (p-Erk), STAT5 (p-STAT5) in mock vector-transduced and F/P-expressing BM-derived MC, in presence or absence of an Akt inhibitor (AKT VIII). β-actin is used as loading control. One representative experiment (out of 3 completely independent experiments) is shown.
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