Skip to main content
. Author manuscript; available in PMC: 2013 Dec 1.
Published in final edited form as: Int J Cancer. 2012 Jun 28;131(11):2693–2703. doi: 10.1002/ijc.27579

Figure 5. FOXO3a regulates sensitivity to the alisertib/ara-C combination.

Figure 5

(a) Alisertib and ara-C cooperate to induce expression of the FOXO3a targets p27 and BIM. MV4-11 cells were treated for 24 hours with 100 nM alisertib, 100 nM ara-C, or both agents. Immunoblotting was utilized to assess the effects of drug treatment on the levels of phospho-FOXO3a, total FOXO3a, p27, and BIM. Tubulin documented equal protein loading. (b) Generation of MV4-11 cells with stable FOXO3a knockdown. Cells were infected with non-targeted control or FOXO3a-targeted shRNA using a lentiviral approach. Stable cell lines were selected with puromycin treatment. Immunoblotting was utilized to assess knockdown efficiency. (c) FOXO3a is required for maximal induction of p27 and BIM by alisertib/ara-C. MV4-11 cells expressing control or FOXO3a-targeted shRNA were treated with 100 nM alisertib, 100 nM ara-C, or both drugs for 24 hours. Protein lysates were subjected to SDS-PAGE and the levels of p27 and BIM were evaluated by immunoblotting. Tubulin documented equal protein loading. (d) Targeted knockdown of FOXO3a blunts the pro-apoptotic effects of the alisertib/ara-C combination. MV4-11 cells expressing control or FOXO3a-targeted shRNA were treated with 100 nM alisertib, 100 nM ara-C, or both drugs for 48 hours. Drug-induced apoptosis was quantified by PI/FACS. n = 3 ± SD, (*p ≤ 0.05, FOXO3a shRNA vs. non-targeted control shRNA).