Extended Data Fig. 7: Analysis of Lip1 Mutants.
Purification of the heterodimeric CerS complex analyzed by SDS-PAGE for A Lip1_F40R and B Lip1_W50A_F51A. Each purifications N = 1. C Proteomic analysis of the purified complex with Lip1_F40R and in D with Lip1_W50A_F51A. Unique peptides of each protein identified are shown in the tables. E Relative CerS enzyme activity measurement in the presence of the fatty-acid-Coenzyme A transport protein Acb1 for the CerS with either WT Lip1, Lip1_F40R or Lip1_W50A_F51A. n = 3 technically independent samples for E and data are presented as mean values ± SD. Exact P-values are shown in Supplementary Table 4. F Levels of phytosphingosine (PHS), in WT cells, lip1Δ cells expressing LIP1_F40R or lip1Δ cells. n = 3 technically independent samples for F and data are presented as mean values ± SD. Exact P-values are shown in Supplementary Table 8. G Tetrad analysis of lip1Δ (orange) cells, expressing LIP1_WT (upper panel, brown), or LIP1_F40R (lower panel, sand).
