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. Author manuscript; available in PMC: 2014 Jan 27.
Published in final edited form as: Clin Cancer Res. 2012 Jul 18;18(18):5020–5030. doi: 10.1158/1078-0432.CCR-12-1072

Figure 3. Blocking AURK gene expression reduces MPNST cell survival in vitro MLN8237 reduces AURKA activity and MPNST cell survival in vitro.

Figure 3

(A) AURKA expression was diminished by AURKA shRNA; AURKB expression was diminished by AURKB shRNA. Blocking each AURK expression at the RNA level was specific as determined by qRT-PCR. (B) Blocking expression of either AURK reduced protein levels of both as detected by Western blot. (C) Phase contrast images of MPNST cells showed a dramatic reduction in cell number after treating with shAURKA or shAURKB. (D) Quantification of cell accumulation by MTS assay showed decreased survival in response to shAURKA and shAURKB. (E) MPNST cells became senescent 48hrs post treatment with MLN8237 as compared to vehicle treated cells as seen in these phase contract images. (F) MLN8237 Aurora kinase A inhibitor reduced survival of all 5 MPNST cell lines tested at an average IC50 of 164nm. (G) Protein levels for AurkA, p-Histone3, total Histone 3, PARP and loading control β-actin for MPNST cells (S462TY) treated with 100nM, 300nM and 1μM of MLN8237 collected at 24hr and 48hr post treatment with drug. Untreated and vehicle treated were used as a negative control and shAURKA treated MPNST cells collected at 4 days post virus treatment serve as a positive control.