Identification of PDZ Domain Containing Proteins Interacting with 1.2 and PMCA4b
Figure 4
Coimmunoprecipitation of PMCA4b. (a) Lysates of transfected HEK 293-PMCA4b cells, kidney, and brain were incubated with monoclonal antibody specific for PMCA4b (for details see Section 2). Protein complexes were then precipitated with protein A/G beads. Western blot of precipitated proteins were probed with a CASK specific antibody. Irrelevant antibodies were used in negative control (α-NFATc2), and transfected HEK 293-PMCA4b cells were used as positive control. (b) Coimmunoprecipitation demonstrated an interaction of PMCA4b and ZO-1. For the IP lysates of transfected HEK 293-PMCA4b cells, HEK 293 cells and ECV cells were precipitated with monoclonal antibody specific for PMCA4b and afterwards IB was performed with antibodies against ZO-1. The negative control was ECV cell lysate incubated with an irrelevant antibody (α-NFATc2) and the positive control was an input of the same cell lysate.
