Fig. 3

Diacylglycerol (DAG) in late Coat Protein I (COPI) vesicle fission. Quantitative data are shown as mean ± s.e.m. Significance was tested using the two-tailed Student’s t test, **P < 0.01, ***P < 0.0001, NS P > 0.05. a Golgi membrane with reduced lipid phosphate phosphatase type 3 (LPP3) level was used for the COPI vesicle reconstitution system; n = 5 independent experiments. b Golgi membrane was isolated from HeLa cells treated with small interfering RNA (siRNA) against LPP3 and then used for incubation in the COPI vesicle reconstitution system. COPI buds on Golgi membrane were then detected by electron microscopy, with representative images shown, bar = 50 nm, n = 3 independent experiments. c Golgi membrane with reduced LPP3 level was used for the COPI vesicle reconstitution system, with rescue of vesicle formation using DAG having varying length of acyl chains as indicated; n = 5 independent experiments. d Golgi membrane with reduced LPP3 level was used for the COPI vesicle reconstitution system, with rescue of vesicle formation using DAG with single unsaturations; n = 5 independent experiments. e Golgi membrane with reduced LPP3 level was used for the COPI vesicle reconstitution system, with rescue of vesicle formation using a polyunsaturated DAG; n = 5 independent experiments. f HeLa cells were treated with siRNA against LPP3 to inhibit COPI transport, with rescue using DAG having varying lengths of acyl chains as indicated. COPI transport in cells was tracked by examining the quantitative colocalization of a COPI-dependent cargo protein (VSVG-KDELR) with Golgi marker (giantin) at different time points as indicated, n = 4 independent experiments. g HeLa cells were treated with siRNA against LPP3 to inhibit COPI transport, with rescue using DAG having single unsaturations. COPI transport in cells was tracked by examining the quantitative colocalization of a COPI-dependent cargo protein (VSVG-KDELR) with Golgi marker (giantin) at different time points as indicated, n = 4 independent experiments. h HeLa cells were treated with siRNA against LPP3 to inhibit COPI transport, with rescue using a polyunsaturated DAG. COPI transport in cells was tracked by examining the quantitative colocalization of a COPI-dependent cargo protein (VSVG-KDELR) with Golgi marker (giantin) at different time points as indicated, n = 4 independent experiments. Source data are provided as a Source Data file