|
|
Blood, Vol. 114, Issue 2, 394-403, July 9, 2009
Survivin mediates aberrant hematopoietic progenitor cell proliferation and acute leukemia in mice induced by internal tandem duplication of Flt3
Blood Fukuda et al.
114: 394
Supplemental materials for: Fukuda et al
Files in this Data Supplement:
- Figure S1. Active caspase-3 expression in Ba/F3 cells expressing wild-type or ITD-Flt3 (JPG, 365 KB)
-
Ba/F3 cells expressing wild-type Flt3 or ITD-Flt3 shown in Fig. 1D were gated based on the side scatter and forward scatter. R1 and R2 represent viable and apoptosing cells, respectively. Active caspase-3 expression in whole cells is shown in the left lower panel and its expression in R1 and R2 are shown in the right panel.
- Figure S2. Effects of survivin deletion on ex vivo cultured HSPC populations (JPG, 126 KB)
-
(A) Fifity thousand bone marrow cells from control survivinflox∕flox and Cre-ER™ survivinflox∕flox mice were plated in 0.3% agar-McCoy5A medium with 15% HI-FBS supplemented with 10 ngs/ml of rmGM-CSF and 50 ngs/ml rmSCF in the presence of 1uM 4OH-Tamoxifen. Colonies in triplicate plates for each of 2 mice were scored on day 7. Data are mean ± SEM from 3 independent experiments. *P<0.05. (B) Mononuclear bone marrow cells from control survivinflox∕flox and Cre-ER™ survivinflox∕flox mice were cultured in 10% HI-FBS/IMDM with 100 ng/ml each of rhFL, rhTpo, and rmSCF in the presence of 1uM 4OH-Tamoxifen for 5 days. At the end of the culture, total cells were counted and stained with APC-anti–c-kit, Streptoavidin-PE–Cy7-biotinylated anti–Sca-1 and PE-conjugated anti–Gr-1, B220, TER119, CD3, and Mac-1 to quantitate KSL cells.
- Figure S3. Effect of in vivo survivin deletion on physiological HSPC populations (JPG, 100 KB)
-
Cre-ER™ survivinflox∕flox or control survivinflox∕flox were treated with 5mg Tamoxifen in corn oil (Sigma) intraperitoneally for 3 consecutive days, allowed to rest for 3 days and subsequently treated for an additional 3 days with Tamoxifen at the same dose to induce survivin gene deletion. Marrow cells were harvested on day 14 after the last dose of Tamoxifen, total nucleated cellularity quantitated and the percentage of reduction in KSL cells and the common myeloid progenitor cells (CMP) in the bone marrow were compared to control survivinflox∕flox mice receiving the same treatment. Data are the mean ± SEM of 3 independent experiments. *P<0.05 compared to control survivinflox∕flox.
- Figure S4. survivin expression RS4;11 and MV4-11 cells (JPG, 39.8 KB)
-
Survivin expression in human acute leukemia cell lines RS4;11 and MV4-11 cultured in 10% FBS plus RPMI-1640 by Western analysis.
|
|
