FIG. 1.

E. histolytica components induce MCP-1 mRNA and protein production from colonic epithelial cells. (A) Confluent Caco-2 and T84 cells were treated with different concentrations of SAP for 8 h. Total cellular RNA was extracted by the TRIzol method, and real-time PCR was performed as described in Materials and Methods. **, P < 0.01; ***, P < 0.001 (over untreated controls). (B) Epithelial cells were treated with 100 μl/mg of SAP for different time periods as indicated. Results in the last lane [Eh (6)] represent results from live E. histolytica amebas (2.5 × 10⁶) in transwells incubated with colonic cell monolayers for 6 h. Data represent changes (n-fold) of mRNA expression over that for the control in experiments repeated thrice with similar results. **, P < 0.01; ***, P < 0.001 (over untreated controls). (C) Caco-2 cells were treated with 100 μl/mg of SAP for different time periods, and whole-cell lysates were subjected to SDS-PAGE and Western blot analysis with MCP-1 antibody and actin antibodies. Ctl, control.