Regulation of EGFR Signaling by miR-203

EGF was from R&D (R&D Systems, MN), and EGFR inhibitor (CI1033 and AG1478) was from Selleck (Selleck, TX). The BD Matrigel™ was purchased from BD Biosciences (BD Biosciences, CA) for invasion assay and tumor cell injections. Precursor for miRNAs (empty vector and miR-203 precursor) and anti-miR inhibitors (control and anti-miR-203) were from GeneCopoeia (GeneCopoeia, MD). RFP reporter vectors were constructed using the Clone-it Enzyme free Lenti-vectors Kit (System Biosciences, CA). Human AREG, EREG, and TGFA full length 3'UTR reporters were constructed using the psiHECKTM-2 vector (Promega, WI). The microRNA binding site mutations were made using the Site-Directed Mutagenesis System kit (Invitrogen, CA). All primers used for these constructs are listed in Supplemental Tables (Table S1). All constructs were verified by DNA sequence analysis.