FIGURE 5.

Binding of DGKη-PH to PI(4,5)P₂-containing liposomes. A, GST-tagged DGKη-PH and GST alone were incubated with liposomes containing PC alone and a 9.5:0.5 molar ratio of PC to PS, PA, and PI(4,5)P₂ as indicated. Liposome-binding proteins were recovered by ultracentrifugation. Liposome binding (precipitate (ppt)) and lack of binding (supernatant (sup)) of GST-DGKη-PH and GST alone were detected by Western blotting with anti-GST antibody. B, the blots were scanned and quantified using ImageJ software. Relative intensities of liposome binding (precipitate) and lack of binding (supernatant) of DGKη-PH and GST alone are indicated, respectively. The data are shown as the means ± S.D. of three independent experiments. Statistical significance was determined using Student's t test versus PI(4,5)P₂ liposome supernatant (**, p < 0.01, ***, p < 0.005) and versus PI(4,5)P₂ liposome precipitate (##, p < 0.01; ###, p < 0.005). Error bars represent S.E.