Figure 2
Alternative translation initiation sites are present in PITX2C mRNA. A) Comparison of the nucleotides flanking the start codon (Met1) and two other in-frame candidate start codons (Met4 and Met35) in the Pitx2c/PITX2C mRNAs from various mammalian species. The Kozak consensus sequence is shown at the top to facilitate the comparison. [GenBank accession nos. for the human: NM_000325, mouse: NM_001042502, rat: AJ222971, cow: NM_001097991, and dog: XM_846277]. B) Schematic representation of the expression constructs designed to study the two PITX2C isoforms. The large and small arrows represent strong and weak AUGSTART codons in each cDNA sequence. C) Immunoblot analysis showing the PITX2C protein isoforms translated from the expression constructs in panel B when transfected into CHO cells. Lysates were run on a 10% Nupage® Novex Bis-Tris gel with MOPS buffer.