High expression of miR-135b and LEMD1 in ALCL. (A-C) Expression of mature miR-135b (A), miR-21 (B), and miR-27a (C) in NPM-ALK (+) ALCL cell lines (Karpas 299, SUDHL-1, and SUP-M2), normal T lymphocytes, and several T-lymphoblastic leukemia cell lines (Jurkat, CCRF-CEM, and Molt4), detected by qRT-PCR analysis. HCT116 colon cancer cells expressing endogenous miR-135b were used as positive control. ND indicates not detected. (D) Attenuation of miR-135b expression by ALK inhibitor WHI-P154. Mature miR-135b in Karpas 299 and SUDHL-1 cells was analyzed by qRT-PCR after WHI-P154 treatment (10μM, 4 hours). (E) Schematic diagram of genomic organization of human LEMD1 gene and miR-135b. As for human LEMD1, several splicing variants have been reported. miR-135b is located in the first intron of LEMD1 longer transcripts. (F) High expression of LEMD1 in ALCL cells, determined as in panel A. (G) Suppression of LEMD1 and pri-miR-135b by WHI-P154 (10μM, 1 hour) in Karpas 299 and SUDHL-1 cells, assessed as in panel D (*P < .05; **P < .01; ***P < .001). (H) Expression of miR-135b in clinical samples of ALCL patients.