Afatinib (BIBW2992) was purchased from Selleck Chemicals (Houston, Texas, USA) as a TKI. DMSO and pyrimethamine were purchased from Sigma Aldrich (St. Louis, Missouri, USA). Afatinib was treated with concentration of 5 mM which obtained over 98% of inhibition of T. gondii growth within the host cells [18 (link)]. Bovine serum albumin was purchased from Bovogen Biologicals (Melbourne, Australia). FITC-conjugated anti-mouse IgG antibody, TRTIC-conjugated anti-rabbit IgG antibody, and horseradish peroxidase-conjugated anti-rabbit or anti-mouse antibodies were purchased from Sigma Aldrich. Antibody against β-actin was purchased from Cell Signaling Technology (Beverly, Massachusetts, USA). Mouse Tg563 monoclonal antibody was cloned in our laboratory. PDCD4 (D29C6) XP® anti-rabbit monoclonal antibody was purchased from Cell Signaling (Boston, Massachusetts, USA) Chemicals for the analytical uses as iodoacetamide was purchased from Sigma-Aldrich. Tris-HCl and urea were obtained from Merck (Branchburg, New Jersey, USA). All other chemicals were acquired from standard sources and were of molecular biology grade or higher.