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. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Mol Genet Metab. 2008 Sep 20;95(3):133–141. doi: 10.1016/j.ymgme.2008.08.003

FIG. 2. Lysosomal ceramide turnover and ceramide content in transduced Farber patient cells.

FIG. 2

FIG. 2

FIG. 2

FIG. 2

Immortalized Farber patient cells were transduced with either oncoretrovirus (RV) or lentivirus (LV) encoding human AC and huCD25. Non-transduced and transduced Farber patient cells were pulsed with [3H-ceramide]-sphingomyelin for 48 h. Lipids were isolated, and then separated by TLC. The AC activity of fibroblasts (A) and B cells (C) are shown. To determine ceramide content, lipid extracts were incubated with E. coli diacylglycerol kinase and [γ32P]ATP. Radioactive ceramide 1-phosphate was isolated by TLC and quantified by liquid scintillation analysis for both fibroblast (B) and B cell (D) extracts. Error bars represent SD; measurements are averages of at least three separate experiments. * p < 0.05, *** p < 0.001, for groups indicated versus non-transduced (NT) controls.