Figure 9. Treatment with Genz-529648 reduces the ceramide content in IB3-1 cells infected with PAO1.

IB3-1 cells subjected to the SL metabolic labeling with (1-³H)sphingosine were treated with [10 µM] amitriptyline alone, in combination with [10 nM] Genz-529648, or with solvent alone and infected with PAO1 as indicated in figure 3. After lipid extraction, (³H)ceramide was separated from the other radioactive SLs by HPTLC, as detailed in the online supplement, and detected by digital autoradiography (total lipid extracts amounts corresponding to 4 µg of cellular proteins were applied on a 4-mm line. Time of acquisition: 48 hours). The digital autoradiography represents data obtained in three different experiments (A). The ceramide content was quantified by specific β-Vision software, and the data reported are the mean ± SE of three independent experiments. Comparisons between groups were made by using Student’s t tests (B).