Figure 2. Expression of ELOVL4ΔC reduces endogenous fatty acid (FA) elongation activities. HEK 293T cells were transfected with a pCE-puro HA-1 (vector), pCE-puro HA-ELOVL4, or pCE-puro HA-ELOVL4ΔC plasmid. Forty-eight hours after transfection, total membrane proteins were prepared from the transfected cells. A: Total membrane proteins (2 μg of protein) were separated by sodium dodecyl sulfate PAGE (SDS–PAGE), followed by immunoblotting with anti-HA antibodies. B: Total membrane proteins (20 μg protein) were incubated with the indicated acyl-CoA (50 μM) and 0.075 μCi [¹⁴C]malonyl-CoA in the presence of 1 mM NADPH, for 30 min at 37 °C. After termination of the reactions, lipids were saponified, acidified, extracted, and separated by normal phase thin layer chromatography (TLC). The radioactivities associated with the reaction product fatty acid (FAs) were quantified using a bioimaging analyzer BAS-2500. Values shown are relative to those for vector-transfected cells, and represent the mean±standard deviation (SD) from three independent experiments. Statistically significant differences compared to vector-transfected cells are indicated (*p<0.05, **p<0.01; t-test). Abbreviations: WT represents wild-type; ΔC represents ELOVL4ΔC; vec represents vector.