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To find out irrespective of whether miRNA are differentially expressed in LSC and HSC, we fractionated 16 AML patient samples and three lineage-depleted (Lin–) cord blood (CB) samples making use of CD34 and CD38 into 4 populations and subjected Just about every 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 diminished in two of 3 samples upon 126KD (Figures 5C and S5D). With each other, these findings suggest that 126KD produces heterogeneous responses with LSC function and frequency reduced in a subset of AML patients.
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(E) Per cent BrdU incorporation into bulk cultures exhibiting proliferation of CTRL and 126OE transduced 8227 cells after a while.
(D) Representative intracellular move plots for that detection of CDK3 and pRB Ser807/811. Graph under signifies 3 unbiased intracellular circulation experiments for every affliction in which the imply fluorescence depth was in contrast. Necessarily mean ± SEM; ∗p=
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In contrast with prior final results displaying miR-126 regulation of ordinary hematopoietic stem mobile (HSC) biking, these purposeful stem consequences are reverse between LSC and HSC. Combined transcriptome and proteome Evaluation demonstrates that miR-126 targets the PI3K/AKT/MTOR signaling pathway, preserving LSC quiescence and selling chemotherapy resistance.