Figure 11

GST-Lepp12 and GST protein preparations used for transfection experiments (see FIGS. 12 and 13) are devoid of measurable endotoxin-like activity, as measured by IL-1beta synthesis by wild-type THP-1 cells. IL-1beta measured by sandwich ELISA in CHAPS lysate of THP-1 cells after 24 h culture of the cells (5 × 10⁴ cells per 200 microL) in the presence of GST-Lepp12 (closed diamonds) or GST (closed squares) was under the sensitivity threshold of the ELISA (10 pg/ml). LPS-induced IL-1beta (open squares) is shown as a positive control. It was above 4 ng/ml when LPS was used at concentrations over 10 ng/ml.