The chimeric firefly luciferase reporter construct of the Men1 CR, 5′-UTR, or 3′-UTR was generated as described (36 (link)). The full-length Men1 CR, 5′-UTR or 3′-UTR was amplified and subcloned into the pmirGLO Dual-Luciferase miRNA Target Expression Vector (Promega, Madison, WI) to generate the pmirGLO-Luc-Menin-CR, pmirGLO-Luc-Menin-5′UTR and pmirGLO-Luc-Menin-3′UTR; cloning was confirmed by DNA sequencing and enzyme digestion. Transient transfections were performed using the Lipofectamine Reagent as recommended by the manufacturer (Invitrogen). The luciferase reporter constructs were transfected into cells along with phRL-null, a Renilla luciferase control reporter vector from Promega, to monitor transfection efficiencies as described previously (38 (link)). Luciferase activity was measured using the Dual Luciferase Assay System, and the level of pmirGLO-Luc-Menin-CR, Menin-5′ or Menin-3′UTR luciferase activity were normalized to Renilla luciferase activity and were further compared with the levels of luciferase mRNA in every experiment. All of the primer sequences for generating these constructs are provided in Supplemental-Table 1. The vector expressing GFP-tagged Menin protein was from OriGene (Rockville, MD).