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Blood, Vol. 113, Issue 8, 1730-1740, February 19, 2009
NOTCH is a key regulator of human T-cell acute leukemia initiating cell activity
Blood Armstrong et al.
113: 1730
Supplemental materials for: Armstrong et al
Files in this Data Supplement:
- Table S1. Characterisics of T-ALL samples at diagnosis (PDF, 29.7 KB)
- Table S2. 250 × 103 T-ALL blasts were cultured with MS5-GFP and MS5-DL1 (± GSI) in individual wells during 30 days (PDF, 11.6 KB) -
Every well was harvested and cells were counted using flow cytometry analysis, with specific morphologic gates for human blast cells. All cells recovered from individual wells were transplanted into a single mouse. Data from 80 mice.
- Figure S1. T-ALL serially engraft NOD-SCID recipients (JPG, 121 KB)
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(A) Phenotype of M22 T-ALL leukemic cells at diagnosis (Patient) and after serial transplantation into NOD-SCID mice. I mouse: primary recipient transplanted with 10E7 cells blasts by intravenous injection. II mouse: secondary recipient; received 10E7 cells from the SPL of the I mouse. III mouse: tertiary recipient; received 10E7 cells from the BM of the secondary mouse. Analysis performed 2 to 3 months after injection when the mice showed signs of sickness. (B) gTCR gene rearrangements in the spleen of the primary (I mouse SPL) and tertiary (III mouse SPL) mouse. shown also are the results obtained from the BM, THY and LN of the same tertiary mouse. Cells at diagnosis (patient) were used as control.
- Figure S2. mRNA levels of c-myc, numb, and spen measured by Q-PCR (JPG, 74.8 KB)
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cDNA were generated from day-3 culture of M18 and M30 T-ALL samples. Cells were co-cultured with MS5-GFP (GFP) oMS5-DL1 (DL1) cells, with (+GSI) or without (−GSI) GSI. Shown are mean values of four measurements normalized using β2microglobulin mRNA levels.
- Figure S3. A minority of cultured cells generated T-ALL in NOD-SCID mice (JPG, 57.9 KB)
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(A) Kinetic analysis of human cell engraftment into NOD-SCID mice. 10E6 T-ALL cells from the spleen of primary mouse, previously injected with cultured M30, were transplanted using IBM injection into 13 mice. Mice were analyzed at 1 (n=5), é (n=5) and 6 (n=2) weeks and % of human CD45+ cells were analyzed by FACS. (B) Cells were phenotyped using human specific monoclonal antibodies. Shown are results obtained from the IBM of a represntative mouse.
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