Figure 2.

SIT4, but not SSO1^ala79, overexpression restores the delivery of FM4-64 to the vacuole in snc cells. (A) Expression of SSO1^ala79 does not restore FM4-64 labeling of the vacuole. snc yeast (JG8) transformed with a plasmid expressing SSO1^ala79 were grown on galactose-containing medium at 26°C, before being shifted to glucose-containing medium for 36 h to deplete Snc1. Cells were incubated with FM4-64 at 26°C and processed for microscopy. (B) SIT4 overexpression restores FM4-64 labeling of the vacuole. snc cells (JG8) transformed with a plasmid expressing SIT4 were grown and labeled with FM4-64 as described above. (C) Ceramine treatment and SIT4 overexpression restore FM4-64 labeling of the vacuole in tlg yeast. tlg1 and tlg2 yeast treated with (+C₂) or without (−C₂) ceramide, or overexpressing SIT4, were labeled with FM4-64 and processed for microscopy.