Figure 1

FOXO4 is a down-stream target of miR-150.

(a) Sequences of the conserved Hsa-miR-150 binding sites in the 3′UTR of wild-type FOXO4 mRNA and a FOXO4 mutant containing seven mutated nucleotides in the 3′UTR of FOXO4 (FOXO4-mut-3′UTR). (b) Luciferase activity assays of the FOXO4-3′UTR or FOXO4-mut-3′UTR reporter in H460 and A375 cells transfected with the pre-miR-150 plasmid or control vector. Experiments were performed in triplicate, and data are expressed as the mean ± SD, **p < 0.01. (c) Western blot analysis showing the alteration of FOXO4 expression in H460 and A549 cells transfected with the pre-miR-150 plasmid, miR-150 inhibitor or their corresponding control vectors. GAPDH served as a loading control. A single band was detected at ~54 kDa, reflecting full-length expression. Fold change in FOXO4 protein level normalized to GAPDH is shown numerically and at the bottom of protein bands.