Fig 2: The in vitro efficacy of AURK-targeting LNA-oligos in ALL and AML cell lines. Cells were incubated with 10 µM of LNA-oligos complementary to AURKA or AURKB or a scrambled control. (a) Microscopic fluorescent imaging of the uptake of a FAM-labeled LNA-oligo by ALL (left) and AML cell lines (right) after 48 h of exposure. Nuclei were stained blue with DAPI. FAM is depicted in green. Bars represent 25 µm. (b) Target-specific reduction of aurora A (left) and aurora B (right) mRNA (upper panels) and protein (lower panels) after exposure of REH cells to an AURKA, AURKB or scrambled control LNA-oligo. Gene expression was determined relative to housekeeping gene RPS20 and was set at 100% for scrambled controls. Brightness and contrast of protein images was optimized and dotted lines indicate where separated samples from the same gel were grouped. (c) Effects of LNA-mediated aurora A or B knockdown on growth of ALL (left) and AML cell lines (right). (d) Effect on cell growth of simultaneous aurora A and B knockdown by LNA-oligos. REH (ALL) and MV4;11 (AML) were incubated for 96 h with 5 µM LNA-oligos targeting AURKA and/or 5 µM LNA-oligos targeting AURKB. Cell growth was determined by trypan blue exclusion and expressed as a percentage of the cell count upon incubation with an equal concentration of a scrambled nontargeting LNA-oligo. AA, LNA-oligo targeting AURKA; AB, LNA-oligo targeting AURKB; scr, scrambled nontargeting LNA-oligo.

Fig 3: Expression patterns of aurora A, B and C in ALL and AML patients and nBM mononuclear cells. Gene expression of AURKA (a), AURKB (b) and AURKC (c) in genetic subtypes of ALL cases (n=297), AML cases (n=237) and nBM samples (n=8) was analyzed by gene-expression array. Asterisks indicate statistical significance with P<0.01 of nBM vs ALL or AML, T-ALL vs precursor B-ALL, and cytogenetic subtypes of precursor B-ALL or AML vs all other subtypes of the B-lymphoid or myeloid lineage. Reverse phase protein arrays (RPPAs) were performed for ALL (n=172) and nBM (n=10) samples to measure aurora A (d) and aurora B (f) protein levels. Values were corrected for total protein and background signals. Statistical significance was determined by comparing groups to all (other) ALL patients. Additionally, protein levels of aurora A (e) and aurora B (g) in ALL (n=10) and AML (n=14) samples were quantified with western blot analysis and corrected for total protein loading by GAPDH expression. Horizontal bars indicate median values. nBM, normal BM mononuclear cells.