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Purposeful validation of sorted fractions was done by xenotransplantation, the result of which was coupled with miRNA expression profiling to generate stem mobile-relevant miRNA expression profiles.
To determine no matter if miRNA are differentially expressed in LSC and HSC, we fractionated 16 AML affected person samples and a few lineage-depleted (Lin–) cord blood (CB) samples applying CD34 and CD38 into four populations and subjected Each individual to world-wide miRNA profiling; the stem mobile content material of every portion was functionally assayed by xenotransplantation (Figures 1A and S1A).
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(J) Proliferation measured by BrdU incorporation assay of CTRL or 126KD transduced 8227 cells in vitro.
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The LSC frequency was lowered in two of three samples upon 126KD (Figures 5C and S5D). Together, these findings counsel that 126KD produces heterogeneous responses with LSC function and frequency reduced inside a subset of AML patients.
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(E) Percent BrdU incorporation into bulk cultures showing proliferation of CTRL and 126OE transduced 8227 cells over time.
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