MN1 suppresses p53 and BIM induction following treatment.

<p>A) Western blot. M1p5 cells expressing MN1 or GFP were exposed to 500 ng/ml doxorubicin for 4 hours as indicated, lysed, and blotted for p53. p53 shRNA denotes M1p5 cells expressing an shRNA targeting p53. B) Western Blot. MF3 cells expressing MN1 or GFP were exposed to doxorubicin as in A, lysed, and blotted for p53 and p21. C) Quantitative RT-PCR assays. M1p5 cells expressing either MN1 or GFP were incubated with 50 ng/ml doxorubicin and 200 nM cytarabine as indicated for 24 hours. Cells were then lysed, total RNA harvested, and RT-QPCR done to determine the relative message levels of the p53 target genes MDM2 and p21. Message levels were set to 1 in untreated GPF- expressing cells. C) Quantitative RT-PCR assays. MF3 cells with or without MN1 overexpression were incubated with the indicated treatment for 24 hours. RT-QPCR was done to determine the relative message levels of BIM. Message levels were set to 1 in untreated GPF-expressing cells. E) Western Blot. M1p5 cells expressing MN1 or GFP were incubated with the indicated therapy for 24 hours, lysed and blotted for BIM protein. Dox20 = doxorubicin 20 ng/ml, Ara100 = cytarabine 100 nM, Ara200 = cytarabine 200 nM.