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Blood, Vol. 112, Issue 12, 4690-4693, December 1, 2008 This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef The PHD fingers of MLL block MLL fusion protein–mediated transformation Blood Muntean et al. 112: 4690 Supplemental materials for: Muntean et al Files in this Data Supplement: Document 1. Supplemental materials and methods (PDF, 69 KB) Figure S1. Detection of MLL fusion constructs by qPCR (JPG, 28 KB) - 293 cells were transiently transfected with MSCV-MLL fusion vectors. RNA was extracted and cDNA generated. qPCR was performed using primers sets specific for the Flag tagged fusion constructs. Expression levels are shown relative to MLL-AF9 expression. Error bars represent standard deviation of experiment performed in triplicate. Figure S2. Expression of MLL-fusion proteins at predicted molecular weights (JPG, 38.1 KB) - 293 cells were transiently transfected and lysed in lysis buffer. Flag tagged MLL fusion proteins were immunoprecipitated with M2 flag beads. Fusion proteins were detected at the predicted molecular weights with anti-Flag antibodies. Figure S3. Co-immunoprecipitation of MLL fusion proteins with Menin (JPG, 34 KB) - 293 cells were transiently transfected and lysed in lysis buffer. Flag tagged MLL fusion proteins were immunoprecipitated with M2 flag beads. Fusion proteins were detected at the predicted molecular weights with anti-Flag antibodies (Top blot) and Menin was detected with the immuno-precipitate (middle blot) and whole cell lysate (bottom blot) with anti-Menin antibodies. Figure S4. MLL-P2-AF9 transforms mouse bone marrow in retroviral transduction and colony assays (JPG, 42.3 KB) - Mouse bone marrow was transduced with MigR1, MSCV, pFMLL-AF9 and pF-MLL-P2-AF9 (which contains only the second PHD finger). Colonies were scored after the 4th round of replating. Similar numbers of colonies were obtained from pFMLL-AF9 and pF-MLL-P2-AF9 transduced bone marrow. Dense, compact colonies were generated from both pFMLL-AF9 and pFMLL-P2-AF9 indicative of transformation. A representation of the MLL-P2-AF9 construct used for the transduction is shown above the bar graph. This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef

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