FIG. 2.

p62 interacts with the aPKC in vivo. (Upper panel) Subconfluent cultures of HeLa cells in 100-mm-diameter plates were transfected with 20 μg of either pCDNA3 (control) or pCDNA3-HA-p62 (p62). Plasmid DNA was removed 4 h later, and the cells were incubated in DMEM containing 10% fetal calf serum for 36 h. Cell extracts (200 μg of protein) were immunoprecipitated with anti-HA antibody. Immunoprecipitates were resolved by SDS-PAGE and immunoblotted with isotype-specific anti-PKC antibodies. The extract (Ext.) lane contains 20 μg of cell protein extract. (Lower panel) Immunoblot analysis of the HA-p62 expressed protein in each assay. Essentially identical results were obtained in two more experiments.