Cx32, SAP97, and CaM coimmunoprecipitation studies using rat Schwannoma cell lysates.
A, lysates of RT4-D6P2T cells were incubated with antibodies directed against SAP97 (SAP97 IP) and probed for Cx32 and SAP97. This co-IP was repeated in the presence of EGTA and Ca2+. B, lysates of RT4-D6P2T cells were incubated with antibodies directed against CaM (CaM IP) and probed for Cx32 and CaM. This co-IP was repeated in the presence of EGTA and Ca2+. C, lysates of RT4-D6P2T cells were incubated with antibodies directed against Cx32 (Cx32 IP) in the presence and absence of λ-phosphatase and probed for Cx32, phospho-Ser, and phospho-Tyr. D, lysates of RT4-D6P2T cells were incubated with antibodies directed against IgG (control IP), SAP97 (SAP97 IP), and CaM (CaM IP) and probed for Cx32, SAP97, and CaM. Input is the RT4-D6P2T lysate. Coimmunoprecipitation experiments were performed in triplicate.