Fig. 2.
Thin-layer chromatography of fluorescently labeled gangliosides reveals accumulation of GM2 ganglioside in the patient's cultured fibroblasts.
Fibroblasts of the patient or a normal healthy control subject were cultured in DMEM containing 1.45 μM of fluorescently labeled BODIPY® FL C5 GM1 ganglioside for 72 h. Total lipids were extracted from cell pellets with 1:1 chloroform/methanol mixture and the gangliosides were separated from neutral lipids and analysed by TLC on silica plates.
(A) The fluorescence image of the TLC plate acquired on G:Box Chemi XQR system. The position of fluorescent GM1 ganglioside used as a standard is shown.
(B) GM1 and GM2 ganglioside bands were quantified in control and patient's fibroblasts by ImageJ software.