siRNA-mediated silencing of TEL-AML1 affects miRNA expression. (A) Schematic representation of the siRNA sequence used to target TEL-AML1 and the localization within the TEL-AML1 translocation breakpoint mRNA (position at bp 15-33, accession no. S78496). A siRNA with no human targets was used as control (see Diakos et al.8 ). (B) Depletion of TEL-AML1 using siRNA. Western analysis was performed from REH cell lysates treated with the control siRNA (left lane) or functional siRNA targeting TEL-AML1 (right lane). TEL-AML1 protein was detected using an anti-TEL antibody from ATLAS and an anti-tubulin antibody for loading control. (C) TEL-AML1 affects miRNA expression. The expression profile of REH cell was analyzed after TEL-AML1 depletion by 2 color miRNA arrays. The fold changes of the 4 most up-regulated as well as those of the 4 most down-regulated miRNAs are shown (mean of 2 experiments). (D) miRNA RT-PCR (Applied Biosystems) was performed to verify the changes in miRNA-494 and miRNA-320a expression upon TEL-AML1 silencing. The data shown depicts the average fold change from 3 independent experiments, using U6 snRNA as a calibration control.
