Interaction of E12 with PIASy.

<p>(A) Coimmunoprecipitation of E12 and PIASy. COS7 cells were transfected with expression plasmids encoding the indicated proteins and incubated for 48 hours. Cell lysates were then prepared, immunoprecipitated (IP) with anti-flag antibody, and analyzed by Western blotting (WB) with anti-myc (top) or anti-flag (bottom) antibodies (Ab). Mouse IgG was used as a control. Input materials (10%) were used as a reference standard. Molecular size markers are indicated on the left. (B) Cell lysates were immunoprecipitated with anti-myc antibody and analyzed by Western blotting (WB) with rabbit anti-flag (top) or anti-myc (bottom) antibodies (Ab). Mouse IgG was used as a control. Input materials (10%) were used as a reference standard. Molecular size markers are indicated on the left. (C and D) Endogenous PIASy and E12 coimmunoprecipitation. Cell lysates from mouse MCs or glomerular lysates from Thy1 rat were immunoprecipitated with anti-PIASy antibody and control IgG. The presence of E12 in the immunoprecipitates was determined by Western blotting with E12 antibody.