In cellulo NDB-GM2 assays of twelve independent treated plates of four sets of human TSD Glial cells.

<p>These sets include, isolated clonal populations highly expressing either the WT alpha subunit (A), the H2 or the H1 beta-alpha hybrid subunit and untransfected (UT) controls (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057908#pone-0057908-t004" target="_blank">Table 4</a> for total MUGS units and cell protein examined in these assays). Loading control (LC) shows the positions of the substrate, NBD-GM2, and its break-down produces; i.e, NBD-GM3, NBD derivative of GM3 ganglioside; NBD-LacCer, NBD derivative of lactosylceramide; NBD-GlcCer, NBD derivative of glcosylceramide; NBD-Cer, NBD derivative of ceramide (note that only low levels of NBD-Cer are seen because of the presence of CBE in the assay, see methods). A) The HPTLC separation of the enriched acidic glycolipid fraction (upper phase Folch extraction) from each plate of cells. B) The HPTLC separation of the enriched neutral glycolipid fraction (lower phase Folch extraction) from each plate of cells. C) Stacked bar graph of the NBD-GM2 hydrolysis products detected in A & B (H2* indicates the moderately high expressing clone, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057908#pone-0057908-t004" target="_blank">Table 4</a>) quantified using the Storm Imager.