Figure 6. Screening hits reverse glycosphingolipid substrate accumulation in H4 cells.

Levels of glucosylsphingosine (GluSph) in frozen cell pellets of HiBiT-GCase-WT and HiBiT-GCase-L444P H4 reporter lines were evaluated by supercritical fluid chromatography (SFC) separation coupled with tandem mass spectrometry (MS/MS) detection and normalized to total cellular inorganic phosphate (Pi) levels. Cells were treated with vehicle (0.3% v/v DMSO) or hit compounds for 3 days (A), except for the cytotoxic hit compound pladienolide B, for which treatment lasted 48 h (B). (Error bars: SD [n = 3 biological replicates]).