Figure 4. HPTLC analysis of the expression of c-series gangliosides in the brains of double-Tg, triple-Tg and WT mice.

Gangliosides were separated by HPTLC using the solvent system of chloroform/methanol/0.2% CaCl₂ (50:45:10, v/v). The bands were visualized by the resorcinol-HCl reagent (A) or immunostaining with anti-A2B5 mAb (B). Each lane contained 3 μg of lipid-bound sialic acid, except lane 4. Lane 1, gangliosides in the brain of WT mice; lane 2, gangliosides in the brain of double-mutant mice; lane 3, gangliosides in the brain of triple-mutant mice; lane 4, authentic GT3 (27 ng). Please note that the amount of GT3 was too low to be detected by the resorcinol-HCl reagent. (C) The concentration of GT3 in the brain of WT, double-mutant and triple-mutant mice. GT3 was quantified by densitometric analysis of (B). n=3. In triple-mutant mice, there was no detectable amount of GT3.