Figure 2.

PKN2 forms complexes with Cdo. (a) Lysates of 293T cells transiently transfected with Cdo, PKN2 or control expression vectors as indicated were immunoprecipitated with an antibody to Cdo. The precipitates and total lysates were immunoblotted with antibodies against Cdo and PKN2, and to α-tubulin as a loading control. (b) Lysates of 293T cells transiently transfected with Cdo, HA-PKN2 or control expression vectors as indicated were immunoprecipitated with an antibody to HA. The precipitates were immunoblotted with antibodies against Cdo or HA. Total lysates were immunoblotted with antibodies to Cdo and PKN2, and to α-tubulin as a loading control. (c) Lysates of C2C12 cells that were proliferating in growth medium (D0) or in DM for the indicated time were immunoprecipitated with antibodies to Cdo and immunoblotted with antibodies to PKN2 or Cdo. Total lysates were also immunoblotted with antibodies to PKN2, Cdo and MHC, and to pan-Cadherin as a loading control. (d) Schematic representation of rat Cdo, the amino acid numbers indicating the deletion constructs. (e) Lysates of 293T cells transiently transfected with Cdo, Cdo deletion mutants, PKN2 or control expression vectors as indicated were immunoprecipitated with PKN2 antibodies and then immunoblotted with antibodies to Cdo. Total lysates were also immunoblotted with antibodies to Cdo or PKN2, and to α-tubulin as a loading control. (f) The schematic representation depicts the domain structure of PKN2 and the deletion of the specific domain. (g) Lysates of 293T cells transiently transfected with GST-tagged PKN2, its deletion mutants, Cdo or control expression vectors as indicated were immunoprecipitated with Cdo antibodies and then immunoblotted with antibodies to Cdo or GST. Total lysates were also immunoblotted with antibodies to Cdo or PKN2, and to α-tubulin as a loading control. Asterisk marks a nonspecific band in the Cdo blot