FIGURE 5.

Isolated nonspecific GPLs (nsGPLs) contain both active and non-active species that differ in their acetylation and methylation patterns. A, TLC of native (N) and deacetylated (DA) nsGPL structural variants. The more apolar nsGPL was named NS2 and the more polar species was named NS3. Upon deacetylation, NS2 increased in polarity, suggesting that it is acetylated in its native conformation. NS3 appeared to be a mixture of two nsGPLs that differ in both their acetylation and methylation patterns. B, GC/MS analyses identified the number of methyl groups located on the sugars of the respective purified deacetylated nsGPLs. Lower panel, TNF-α production was measured upon exposure to total lipid (TL) and native and deacetylated (nsGPLs). NT, not-treated. n = 3 separate experiments ± S.E.