The miR 150/NOTCH3 interaction is operative in T-ALL cell lines. (A) RT-PCR to detect miR-150 in the T-ALL cell lines Jurkat, MOLT-3, and DND41 72 hours after transduction with the lentiviral vector LV-pre-miR-150 or LV-control. The level of expression was normalized to that of the small noncoding RNA RNU44. The figure shows the average of 3 independent experiments plus or minus SD. (B) Western blot showing that miR-150 overexpression induces a decrease in the levels of endogenous full-length (FL) NOTCH3 protein in the T-ALL cell lines. α-tubulin was used as a loading control. The numbers below the bands indicate the relative ratio between the NOTCH3 and the α-tubulin bands, normalized by setting the value obtained for the controls at 1. (C) Western blot showing that miRZip anti-miR-150 vector (LV-miRZip-150) prevents attenuation of NOTCH3 protein levels in DND41 cells cotransduced with LV-pre-miR-150. β-actin was used as a loading control. The numbers below the bands indicate the relative ratio between the NOTCH3 and the β-actin bands, normalized by setting the value obtained for the untreated cells at 1. (D) RT-PCR to detect NOTCH3 mRNA in the T-ALL cell lines Jurkat, MOLT-3, and DND41 72 hours after transduction with the lentiviral vector LV-pre-miR-150 or LV-control. The level of expression was normalized to that of the housekeeping gene β2-microglobulin. The figure shows the average of 3 independent experiments plus or minus SD. *P < .05 (t test). **P < .001 (t test).