Comparison of bSMase treatment effects on backbone fluorophores in different SM derivatives. HeLa cells were incubated with TFSM 1 , the FRET probe, or BODIPY-FL-C 12 -SM. Molecules were removed, and cells were bSMase-treated and detached. Cells incubated with TFSM 1 were fixed stained with BODIPY-FL-DBCO, and backbone fluorophores were analyzed by flow cytometry (BODIPY-FL for TFSM 1 ; BODIPY-FL-C 12 -SM and BODIPY-TR for the FRET probe). Mean fluorescence in bSMase-treated samples was normalized to untreated controls (100%). n = 3.
