CGI-58 affect C20PUFA abundance in the lipid droplet membrane.

<p><b>(A)-(D)</b> Loss of <i>cgi-58</i> in AMPK mutant (<b>A</b> and <b>B</b>) and control <i>daf-2</i> (<b>C</b> and <b>D</b>) dauer larvae led to a lipid droplet-specific reduction of C20PUFAs. LD = lipid droplets. Cyto = cytoplasm. Purified lipid droplets and remaining cytoplasm were subjected to fatty acid methyl ester (FAME) extraction followed by GC/MS analysis. Short/branched fatty acids are the combined total percentage of C14:0, C15ISO, C16ISO, C16:0, C17ISO, C16:1n7 and C17Δ. C18 fatty acids are the combined total percentage of C18:0, C18:1n11, C18:1n10, C18:1n9, C18:1n7, C18:2n6 and C18:3n6. C20PUFAs are total percentage of C20:3n6, C20:4n6, C20:4n3 and C20:5n3. ** indicates statistical significance (P<0.01) and error bars indicate SD of three independent experiments in <b>(A)-(I)</b>. <b>(E)-(F)</b> The amount of all major C20PUFAs were decreased in the lipid droplets of <i>daf-2; aak(0); cgi-58</i> <b>(E)</b> and <i>daf-2; cgi-58</i> <b>(F)</b> dauer larvae. <b>(G)</b> Long chain fatty acids were decreased in <i>CGI-58</i> MEF mutants. Total cell lysates were subjected to FAME extraction followed by GC/MS analysis. Long chain fatty acids include fatty acids with a carbon chain longer than 20 and certain aromatic fatty acids. <b>(H)</b> C20PUFAs were lost from the phospholipid portion of the lipid droplets of <i>daf-2; aak(0); cgi-58</i> dauer larvae. PL = phospholipid. <b>(I)</b> Some C18 fatty acids were completely absent in the triglyceride portion of the lipid droplets of <i>daf-2; aak(0); cgi-58</i> dauer larvae. TG = triglyceride.