Expression of PNT in E. histolytica. (A) Immunoblot analysis of native EhPNT. Approximately 10 μg of total lysate was electrophoresed on a 12% SDS-polyacrylamide gel and subjected to an immunoblot assay with anti-EhPNT antibody or preimmune serum. M, molecular mass marker. (B) Immunoprecipitation of EhPNT. The lysates derived from the transformant expressing either EhPNT-HA (“PNT-HA”) or EhCpn60-HA (“CPN60-HA”) and the wild-type strain (“WT”) were subjected to immunoprecipitation with anti-HA antibody, followed by immunoblot analysis with anti-HA antibody. Lysate derived from the wild-type amoebae was also used directly for immunoblot analysis as a control. An arrowhead and an arrow indicate heavy and light chains of anti-HA antibody, respectively. IP, immunoprecipitation; IB, immunoblot. (C) Effect of heat treatment on the mobility of EhPNT on an SDS-PAGE gel. Approximately 10 μg of total lysate was electrophoresed on a 12% SDS-polyacrylamide gel and subjected to an immunoblot assay with anti-EhPNT antibody. M, molecular mass marker.