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. 2016 Oct 5;6:34648. doi: 10.1038/srep34648

Figure 1. TALDO1 has two translation initiation sites.

Figure 1

(A) TALDO1 isoforms were analysed by western blotting in NIH/3T3 (wild-type and TALDO1 knockout [KO]) and human cell lines (HepG2, Hep3B, HLE, HLF, HuCCT1, and HuH28 cells). Three and a half micrograms (NIH/3T3) or 27.6 μg (human cell lines) of total protein was loaded in each lane for sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Actin was used as a loading control. Immunofluorescence images of wild-type NIH/3T3 and TALDO1 KO cells stained with anti-TALDO1 antibody. Cell nuclei were stained using DAPI. (B) Schematic representation of the conserved Mus musculus (mouse) Taldo1 sequence around two translation initiation sites. Two potential in-frame translation initiation sites, designated sites A and B, are present in the Taldo1 mRNA. Underlined nucleotides indicated potential translation initiation AUG-sites and critical nucleotides at positions −3 and +4 according to the optimal Kozak translation initiation consensus sequence. (C) Comparison of the region of Taldo1 sequence around two translation initiation sites from Homo sapiens (human), Gallus gallus (chicken), Danio rerio (zebrafish), Drosophila melanogaster (fruitfly), Caenorhabditis elegans (nematode), and Saccharomyces cerevisiae (yeast). (D) Schematic representations of the native and mutant 5′ untranslated regions (UTRs) of the Taldo1 gene used in this study. (E) Western blot using anti-HA antibody showing the expression of native 5′ UTR-TALDO1-3×HA, M1A-3×HA, S2A-3×HA, M11A-3×HA, 3×HA-TALDO1L, and 3×HA-TALDO1S transfected into wild-type NIH/3T3 cells.