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Diagnosis of a variant of APL was made according to the morphology, fluorescence in situ hybridization (FISH), and flow cytometric analysis [human leukocyte antigen DR (HLA-DR Web site; see the Supplemental Materials link at the top of the online article).
Then, tamibarotene (6 mg/kg) was given, which mildly reduced the leukemia burden.These results highlight essential features of pathogenesis in APL in more detail.BCOR appears to be involved not only in human congenital diseases, but also in a human cancer.Thirty-five months after diagnosis, the patient relapsed with gradually reducing blood counts and overt coagulopathy.A bone marrow sample showed that 90% of the cells were leukemic blasts with the recurring chromosomal translocation, FISH abnormality of , and aberrant expression of CD56 and CD13.Clinical samples were collected under institutional review board–approved protocols and with written informed consent in accordance with the Declaration of Helsinki.
Briefly, total RNA from bone marrow nuclear cells before treatment was isolated with an RNeasy plus mini kit (QIAGEN).
(See Figure 3A for composition of the deletion mutants.) For coimmunoprecipitation, expression plasmids of Myc- or FLAG-tagged proteins were transiently transfected with Lipofectamine 2000 (Invitrogen) into 293T.
After 48 hours of incubation, cells were harvested, washed once in phosphate-buffered saline and lysed in 150m M Na Cl, 50m M Tris [tris(hydroxymethyl)aminomethane]-HCl (p H 7.4), 1m M EDTA (ethylenedimainetetraacetic acid), 1% Triton X-100, and Phos STOP (Roche) on ice for 20 minutes.
Annealing steps were 70°C for 30 seconds from the 6th to 10th cycles and 68°C for 30 seconds from the 11th to 30th cycles.
were amplified from total RNA of bone marrow samples with Phusion DNA polymerase (Finnzymes).
All PCR reactions were accomplished under the following conditions.