FIG 3.

Nitric oxide regulates the amount of Ehmeth-enolase inhibitory complex formed. (A) Ehmeth samples from nuclear lysates of pJST4-Ehmeth and pJST4 Ehmeth E. histolytica trophozoites that were treated with 350 μM GSNO for 1 h was immunoprecipitated (IP) with a monoclonal anti-HA (α HA) antibody. The presence of enolase among the immunoprecipitated proteins was detected by Western blotting with an enolase antibody (left). The presence of CHH-tagged Ehmeth among the immunoprecipitated proteins was detected by Western blotting by using a histidine antibody. The immunoprecipitation experiments were also performed with nuclear lysates of pcontrol E. histolytica trophozoites as a negative control for the expression of CHH-tagged Ehmeth and for the immunoprecipitation of enolase (right). (B) Western blot analysis of nuclear proteins prepared from GSNO-treated pJST4-Ehmeth and pJST4-Ehmeth E. histolytica trophozoites. The proteins were separated on 12% SDS-PAGE gels and analyzed by Western blotting with an HA antibody, an enolase antibody, or an actin antibody. The figure displays a representative result from at least three independent experiments.