Fig 8. Microsomal GlcN-(acyl)PI synthesis activity in flc mutants is increased.
Microsomes were produced from WT or 1Δ2Δ3tyΔ cells grown for 12 h in the presence of Doxy and sorbitol, and were labeled using 80 nM UDP-[3H]-GlcNAc (4 μCi), 1 mM ATP and 1 mM CoA. In the last lane, to produce GPI intermediates without the acyl chain bound to the inositol moiety, CoA and ATP were omitted and 1 mM G3P was added in order to deplete microsomes of preexisting acyl-CoA. Lipids were extracted and separated by TLC.