Decreased expression of miR-150 in T-cell lymphomas. (A) Northern blot analysis of miR-150 in T-cell lymphoma/leukemia cell lines. Fold changes in miRNA levels were determined by densitometry and are shown below the gels after normalization to the level of 5S tRNA in normal CD4⁺ T cells (CD4-1), which were assigned a value of 1.00. Northern blots of miR-16-1, miR-29a, miR-29b, and U47 (another control for 5S tRNA) are also shown. Cell sorter was used to purify CD4⁺ T cells from lymph nodes or peripheral blood collected from primary samples (>95% purification). (B) Northern analysis of miR-150 in CD4⁺ T cells (n = 2) and primary samples of PTCL (2 cases of PTCL NOS), and 4 CTCL; 5S served as a control. Northern blots of miR-16-1, miR-29a, miR-29b, and U47 are also shown. (C) qPCR analysis of miR-150. (Left) qPCR analysis of miR-150 in normal CD3⁺ T cells (n = 15), CD4⁺ T cells (n = 14), CD8⁺ T cells (n = 8), and CD56⁺ T/NK cells (n = 11). (Right) qPCR analysis of miR-150 expression in 49 samples of CD4⁺ T-cell lymphoma, including CTCL (n = 12), AITL (n = 10), ATLL (n = 7), and PTCL NOS (n = 20). Y-axis: 2^-ΔCt values for miRNA expression. Expression levels were normalized by data from PTCL2, which was assigned a value of 1.0; qPCR was conducted in duplicate, yielding similar results. P values were calculated using Mann-Whitney U test. Asterisks (*) indicate statistical significance: ***P < .001.