FIGURE 2.

Lipid binding activities of GST-tagged PHs of DGKδ, -η, and -κ. A and C, equimolar amounts (100 pmol) of PI, PI(3)P, PI(4)P, PI(5)P, PC, PI(3,4)P₂, PI(4,5)P₂, PI(3,4,5)P₃, PA, and PS (A) and PA, PS, CL, PC, SM, Sph, PG, PE, PI, and C1P (C) were spotted onto nitrocellulose membranes as indicated, respectively. The membranes were incubated with GST-tagged PHs of DGKδ, -η, and -κ as indicated. The lipid-bound PHs were detected by immunostaining with anti-GST antibody. B and D, the blots were scanned and quantified using ImageJ software. PI(4,5)P₂ (B) and PA (D) binding levels of DGKη-PH were set to 100%, respectively. The data are shown as the means ± S.D. of three independent experiments. Statistical significance compared with the PI(4,5)P₂ (B) and PA (D) binding activities of DGKη-PH (* and ***) or DGKδ-PH (# and ###) was determined using Student's t test (*, p < 0.05; ***, p < 0.005; #, p < 0.05; ###, p < 0.005). Error bars represent S.E.