Fig. 4: Dihydroceramides (DH-Cer) induce fat clearance.

A Representative photomicrographs of T6PNE cells treated for 2 days with 0.25 mM palmitate and the indicated compounds, followed by staining for fat with Nile Red. B Quantification of Nile Red staining from A. C Inhibition by siRNA to SPNS2 or S1PR3 of fat clearance induced by DH-Cer. T6PNE cells were transfected with siRNAs to SPNS2 or S1PR3. Two days later, DH-Cer was added for 2 days, followed by staining with Oil Red O. D Quantification of the number of cells positive for Nile red from C, demonstrating that DH-Cer-induced fat clearance requires SPNS2 and S1PR3. E DES-1 inhibitors GT-11and B-0027 increase fat clearance. T6PNE cells were treated with 0.25 mM palmitate and DMSO or NCT. F Quantification of the Nile Red cells shown in E. Values represent the mean ± SE of 6 biological replicates, *p < 0.05 (vs DMSO or scrambled siRNA). Scale bar = 100 μm.