miR-125b upregulates VEGFA expression in part through suppression of TET2. (A) TET2 messenger RNA (mRNA) levels were assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in AF9+Ri125b+Dox and AF9+Ri125b−Dox cells at day 6. 18S ribosomal RNA (rRNA) was used as an internal control (n = 3). (B) shRNA against TET2 or a control shRNA was transduced into primary MLL-AF9–only AML cells. TET2 expression levels were determined by qRT-PCR 7 days after transduction (n = 3). (C) MLL-AF9–only leukemia cells were infected with shTET2, shCtrl, or a mock infection. After 7 days of puromycin selection, total mRNAs were harvested for TET2 expression analysis. 18S rRNA was used as an internal control (n = 3). (D) VEGFA mRNA levels were analyzed in MLL-AF9–only AML cells that were infected with shp53 or a control shScr vector. 18s was used as an internal control (n = 3). (E) TET2 cDNA or a control vector was transduced into AF9+Ri125b AML cells in the presence or absence of Dox treatment. Transduced cells were fluorescence-activated cell sorted 36 hours post infection, and the levels of VEGFA expression were determined by qRT-PCR and normalized to 18S rRNA (n = 3; error bars represent standard deviations; *P < .01; **P < .001 in all panels). NS, not significant.