FIGURE 1.

AKR1B10 association with HSP90 α. A, coimmunoprecipitation. AKR1B10 protein complexes in HCT-8 cells were immunoprecipitated with a specific anti-AKR1B10 antibody and separated in 12% SDS-PAGE. i, Coomassie Blue staining showing a candidate protein at ∼90 kDa (arrow). The numbers on the left show the molecular weight (in kilodalton) of protein markers. ACCA, acetyl-CoA carboxylase-α. ii, matrix-assisted laser desorption ionization mass spectrometry of the candidate protein. The image shows the identified peptide that is identical to residues 101–112 (ADLINNLGTIAK) of HSP90α. B, Western blot analysis. Immunoprecipitated AKR1B10 complexes were subjected to Western blot analysis as described under “Materials and Methods” to confirm the presence of HSP90α. Lane 1, rabbit IgG negative control. Lane 2, immunoprecipitates with anti-AKR1B10 antibody (α-B10). Lane 3, 20% input. C, HSP90α pull-down carried out with purified HSP90α protein as described under “Materials and Methods.”