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. 2012 Mar 12;7(3):e33140. doi: 10.1371/journal.pone.0033140

Figure 6. The p58 isoform of IGF2BP2 is not generated by protease cleavage.

Figure 6

A. A schematic representation of the frameshift mutation. A single nucleotide deletion at position 198 resulted in a premature stop codon in the ORF starting at the double Met but did not affect the downstream ORF starting from the internal Met69. A Kozak sequence was also added upstream of the canonical double Met. B. Western blot analysis of the frameshift construct expressed in 3T3-L1 preadipocytes. Empty vector (pBabe) or the frameshift construct (Δnt198) was introduced into the cells using retrovirus. Cellular proteins (10 µg/lane) were resolved by 10% SDS-PAGE and blots were probed with antibody to IGF2BP2 C-terminal peptide (CRB Ltd) and with antibody to the p85 subunit of PI3-kinase as a loading control. Positions of molecular weight markers are indicated to the right.