Fig. 6.

Substrate concentration-dependence. (A) Various concentrations of [¹⁴C]SAG were incubated with the partially purified rat brain DG lipase from the CHT 5-I column (15 µg per assay). Except for the substrate concentrations, the assay conditions were the same as those for the standard assay. (B) Various concentrations of [¹⁴C]SAG were incubated with HA-rDDHD2 (closed circles) prepared using anti-rat DDHD2 antibody. Control (closed triangles) means the DG lipase activity using the same aliquot of protein sample obtained from vector transfected cells. (C) Control sample and HA-rDDHD2 sample were analysed by western blotting using anti-rat DDHD2 antibody and SYPRO Ruby staining following SDS-PAGE. The rectangular area in SYPRO Ruby staining is magnified in the right panel. Data are the means of triplicate measurements. Error bars represent standard errors of the mean. Data are representative of three independent experiments.