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Blood, Vol. 108, Issue 8, 2770-2779, October 15, 2006
JAK2T875N is a novel activating mutation that results in myeloproliferative disease with features of megakaryoblastic leukemia in a murine bone marrow transplantation model
Blood Mercher et al.
108: 2770
Supplemental materials for: Mercher et al
Files in this Data Supplement:
- Table S1. Phosphopeptides identified by LC-MS/MS in CHRF-288-11 cells (PDF, 26.9 KB) -
Tandem mass spectra were collected in a data-dependent manner with an LCQ ion trap mass spectrometer (ThermoFinnigan), using a top-ten method, a dynamic exclusion repeat count of 1, and a repeat duration of 10 sec. TurboSequest (ThermoFinnigan) searches were done against the NCBI human database released on February 23, 2004 containing 27,175 proteins allowing for tyrosine phosphorylation (Y+80) and oxidized methionine (M+16) as dynamic modifications. Sequence assignments were accepted on the score-filtering criteria described previously,26 with the exception that assignments having score characteristics of false-positive assignments, determined by searching against a database of reversed protein sequences,47 were not accepted.
§ indicates known phosphorylation site.
- Figure S1. Studies of Jak2, STAT5, ERK1/2 and AKT phosphorylation in Ba/F3 EpoR cells over-expressing either JAK2WT, JAK2T875N or JAK2V617F (PDF, 270 KB) -
Ba/F3 EpoR cells were transduced with each of the constructs, sorted for GFP after 24 hours, grown in presence of IL3 for 8 days and treated as in Figure 3 prior to immunoprecipitation and western blot analysis. No significant difference in the phosphorylation levels of Jak2, STAT5, ERK1/2 or AKT was observed between JAK2T875N and JAK2V617F expressing cells.
- Figure S2. Kinetics of WBC and hematocrit percentages in C57Bl/6 and Balb/c transplanted with WT and T875N Jak2 (PDF, 95.3 KB) -
Mean value from at least 5 animals +/- s.e.m is represented.
- Figure S3. Levels of EPO in plasma are not elevated in Jak2T875N animals (PDF, 73.5 KB) -
Plasma levels of EPO were measured by ELISA. Animals treated with 5-fluorouracil 5 days before blood collection showing severe anemia were used as positive controls. Bottom graph is a close-up of the top graph to better show the low EPO concentration samples.
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