Figure 5. 32D-IRS1 cells express higher levels of Annexin A2.
A. 32D cells lacking or expressing IRS1 were treated with IL-3 for 30 minutes. Cell lysates were prepared and loaded on a gel, separated by SDS-PAGE, and stained with Imperial™ protein stain overnight. The region of the gel corresponding to ~38kDa was excised for both samples. Proteins were eluted and analyzed by tandem mass spectrometry. Peptide representation was compared between the 2 samples. B. 32D cells lacking or expressing IRS1 and/or IRS2 were treated in the presence or absence of IL-3, supplied as 5% WEHI-3B supernatant which contains 0.75% serum. Cell lysates were prepared and the western blot was probed with anti Annexin A2. Densitometry was used to quantify bands. C. Amino acid sequences for mouse Annexin A2 (NP_031611.1) and mouse BAK (CAA73684.1) were aligned using the NCBI site. Annexin A2 contains 339 amino acids and BAK contains 208 amino acids.