Figure 9.

[¹⁴C]Acetate labeling of phospholipids and neutral lipids in At1g52760-overexpressing yeast cells. The peanut MGAT homolog of Arabidopsis At1g52760 was overexpressed in wild-type yeast in the presence of [¹⁴C]acetate (0.5 μCi mL⁻¹). The yeast cells were grown in SM-U containing 2% Gal for 24 h, and equal absorbance (A₆₀₀ = 20) of cells was harvested and lipids were extracted. A, Incorporation of [¹⁴C]acetate into neutral lipids. Lipids were separated on TLC using petroleum ether:diethyl ether:acetic acid (70:30:1, v/v). 0, Origin; 1, LPC; 2, PI, phosphatidylinositol; 3, PS, phosphatidylserine; 5, PC, phosphatidylcholine; 6, PE, phosphatidylethanolamine; 4, 7, 8, and 9, unknown; NL, neutral lipid. B, [¹⁴C]Acetate incorporation into yeast phospholipids. Two-dimensional TLC using chloroform:methanol:ammonia (65:25:5, v/v) was used in the first dimension followed by chloroform:methanol:acetone:acetic acid:water (50:10:20:15:5, v/v) in the second dimension. Values are means ± sd of three independent experiments. FFA, Free fatty acid; OD, optical density.