Figure 2. TSP-1 or NTSP pulls down surface TcCRT.

2A, TSP-1 or NTSP pulls down a metabolically labeled T. cruzi 48 kDa protein whereas BSA control does not. Extracted ³⁵S-methionine labeled trypomastigote membrane fraction was incubated with NTSP, TSP-1 or BSA coated Dynabeads to pull down interacting proteins. The Pull downs were separated by SDS polyacrylamide gels, transferred onto nitrocellulose (NC) membranes and developed by autoradiography. 2B, NTSP or TSP-1 pulls down double labeled T. cruzi 48 kDa surface protein whereas BSA control does not. Extracted ³⁵S-methionine and NHS-biotin double labeled surface proteins of T. cruzi interacting with NTSP, TSP-1 or BSA coated Dynabeads were recruited with streptavidin Dynabeads, separated by SDS polyacrylamide gel, transferred onto NC membranes and developed by autoradiography. 2C, Purification of rTcCRT. Coommassie blue stain of two eluted fractions of affinity purified His-tagged rTcCRT. 2D, TSP-1 or NTSP pulls down a TcCRT as evidenced by immunoblotting using anti-TcCRT IgG. 2E, Blot of figure 2D was stripped and reprobed with an isotype control antibody. This figure shows the results from a representative experiment of three independent experiments performed with similar results.