FIGURE 4.

Scx physically associated with E12. A, co-immunoprecipitation of FLAG-Scx and Myc-E12 from co-transfected CHOK1 cells lysate was detected by Western blot. IP, immunoprecipitation. IB, immunoblot. B, immunofluorescence of AGE-treated MCs with antibodies against the Scx (green) and E12 (red) protein showing the localization of the two proteins in the nucleus. White arrows indicated partial co-localization of Scx and E12 proteins. The cells were counterstained with DAPI to visualize the nuclei. C, Western blot of immunoprecipitates from AGE-treated MCs lysates using a polyclonal anti-E12 antibody or control IgG. The reaction mixture was loaded as input. D, kidney lysates from diabetic or nondiabetic (Control) mice were immunoprecipitated (IP) with anti-E12 antibody and control IgG. The presence of Scx in the immunoprecipitates was determined by Western blot with anti-Scx antibody. The control antibody does not immunoprecipitate Scx. Aliquots of the lysates from diabetic kidney were loaded as positive controls (PC). One of five independent experiments is shown in each panel.