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Figure 3


Fig. 3. Knockdown of mLgl increases interaction of aPKC-PAR-6 with PAR-3 or Cdc42. (A) Confluent monolayer of control cells (21-1 or 21-7) or mLgl kd cells (24-15 or 24-16) was lysed with lysis buffer and clarified by centrifugation. The supernatant fraction (Sup, lower panels) containing total 1 mg protein was subjected to immunoprecipitation with anti-PAR-6ß antibody and precipitated proteins were analyzed by western blot (IP, upper panels). Three independent experiments were performed for each clone and numbered as 1, 2 and 3. Notice that the co-precipitation of PAR-3 or Cdc42 with PAR-6 was increased in mLgl kd cells. (B-D) Quantification of the amount of PAR-3 (B), aPKC{lambda} (C) or PAR-6ß in IP fraction. The amount of PAR-3 increased significantly in both mLgl kd cells (*P<0.005, **P<0.001), whereas the amount of aPKC{lambda} or PAR-6ß was not changed in these cells.