Fig. 1.

Deletion of EMP24 partially rescues the lcb1-100 phenotype. A: Pulse-chase analysis of the maturation of the GPI-anchored protein, Gas1p, in wild-type RH2888 (Wt), lcb1-100 cells, RH3948 (lcb1), emp24Δ cells, RH3950 (emp24Δ), and double mutant cells RH3949 (emp24Δ lcb1-100). The precursor form (p) of Gas1p is generated in the ER and maturation (m) occurs upon arrival in the Golgi compartment. The percentage of mature form is quantified. B: Serial dilutions of wild-type and mutant cells were spotted on YPD plates and incubated at the indicated temperatures for 3 days. Quantification by ESI-MS of the major ceramide and IPC species produced in each strain after incubation overnight at 30°C with or without a 1 h shift to 37°C before harvesting. Asterisks indicate P < 0.05 for the pairwise combinations designated. C: Dilution series of strains were spotted as in (B) onto plates containing 0.5µg/ml of myriocin, grown at 30°C. Plates shown are representative of at least three replicates per condition.