Fig. 3.

TLC analysis of various recombinant acyltransferase acylation products using 1-MAkG as substrate and their relative protein expressions. Aliquots of 30 μg microsomal membranes with recombinant human acyltransferase expression, naïve Sf9 or High5 microsomal membranes were used for acyltransferase reactions using 0.1 mM 1-MAkG and 0.05 mM [¹⁴C]oleoyl-CoA (specific activity at 2 μCi/μmol) in 150 mM KHxPO4 (pH 7.4) buffer at 37°C for 15 min, as described in the Materials and Methods, and the resulting enzyme products were analyzed by TLC. Aliquots of 1 μg acyltransferase microsomal membranes were subjected to quantitative immunoblot analysis, as described in the Materials and Methods. The relative protein expression level of each acyltransferase was reflected by the Flag band intensity and normalized against that of DGAT1, which was designated as 1 protein unit. A: TLC profile of lipid products. *Denotes fatty acid methyl ester (FAME), a nonspecific product formed during lipid extraction and analysis. B: Anti-Flag immunoblot for various acyltransferases.