Table 1

Hydrolysis of BODIPY-LacCer in normal vs. ML-IV fibroblasts

Incubation conditions% hydrolysis
NormalML-IV
Pulse (1 hr; 19°C) → Chase (1 hr; 37°C)76
Pulse (1 hr; 37°C) → Chase (1 hr; 37°C)4232
Pulse (1 hr; 37°C) → Chase (3 hr; 37°C)7468
  • Cells were incubated with C5-DMB-LacCer/BSA for 1 hr at 19°C or 37°C, washed, and further incubated for 1 or 3 hr at 37°C before lipid extraction and analysis. Percent hydrolysis is [(C5-DMB-metabolites)/(C5DMB-LacCer + C5-DMG-metabolites)] × 100, where the C5-DMB-metabolites were C5-DMB-Cer, -GlcCer, and -SM. For a 1-hr pulse/1-hr chase (both at 37°C), the values in normal vs. ML-IV cells, respectively, were C5-DMB-LacCer (58.2 ± 1.1% vs. 68.2 ± 0.4%), -Cer (16 ± 0.6% vs. 11.6 ± 0.2%), -GlcCer (12.3 ± 0.2% vs. 9.9 ± 0.1%), and -SM (13.3 ± 0.2% vs. 10.3 ± 0.1%). GlcCer, glucosylceramide; SM, sphingomyelin.