Fig. 1
From: Ubiquitous Induction of p53 in Tumor Cells by Antisense Inhibition of MDM2 Expression
Characterization of MDM2 antisense oligonucleotides.
(A) Sequence of MDM2 antisense phosphorothioate oligonucleotides. The top sequence represents the noncoding strand of human MDM2. The numbers indicate distance from the translation initiation codon. (B) Screening of MDM2 antisense oligonucleotides. MCF-7 cells stably transfected with the BP100-luc reporter were treated with 50 nM of MDM2 oligonucleotides for 20 hr. p53 transcriptional activity was determined as luciferase activity/unit protein. M4 is a 4-bp mismatch control of AS. LC, control with lipofection alone. (C) Activation of p53 by AS2 is sequence-specific. JAR cells stably transfected with the BP100-luc reporter were treated with AS2, mismatch control oligonucleotides of AS2 (AS2M2, 2-bp mismatch; AS2M4, 4-bp mismatch), and an unrelated oligonucleotide K. (D) Inhibition of MDM2 expression by AS2. MDM2 levels in JAR cells treated for 20 hr with AS2 or AS2M4 control oligonucleotide were determined by Western blotting using monoclonal antibody 3G9. JAR cells treated with lipofectin alone, loaded at 1-, 2-, and 4-fold dilutions, were used as controls.