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. 2009 May 8;284(28):18863–18872. doi: 10.1074/jbc.M109.008623

FIGURE 4.

FIGURE 4.

Binding of GST-CPE to Cld3-transfected HEK cells. Three days after transfection, cells were incubated with 1 μg/ml GST-CPE-(116–319) (left, middle) or GST-CPE-(194–319) (right) for 1 h at 37 °C, washed with PBS, and the amount of bound GST-CPE and claudin in the lysate analyzed by SDS-PAGE and Western blot using anti-GST (top) and anti-Cld3 (bottom) antibodies. Left, GST-CPE-(116–319) bound to Cld3wt-transfected (Cld3) but not to non-transfected (n.t.) cells. Middle, GST-CPE-(116–319) bound to Cld3-N148D and Cld3-L150A to a similar extent as to Cld3wt. In contrast, binding to Cld3-N148D/L150A was strongly reduced. Right, similar results were obtained for GST-CPE-(194–319). Representative blots are shown.