Fig. 3.
Identification of the 22-kDa URE3-BP-interacting protein as EhC2A. (A) Western blots were performed on amebic whole-cell lysates or recombinant purified His6-tagged EhC2A using anti-EhC2A rabbit serum developed with recombinant purified GST-tagged EhC2A. Preimmune serum lanes are indicated. α, anti. (B) Trophozoites were lysed in the presence of 5 mM EGTA (E) and immunoprecipitated (IP) with anti-EhC2A rabbit serum (EhC2A) or control rabbit serum (Control). Coimmunoprecipitated proteins were detected by Western blotting (WB). (C) Trophozoites were lysed in the presence of 5 mM EGTA (E) or 0.1 mM CaCl2 (C) and immunoprecipitated (IP) with anti-URE3-BP MAb 4D6 (URE3-BP) or control mouse MAb (Control). Coimmunoprecipitated proteins were detected by Western blotting (WB).