Figure 1.

Delivery of FM4-64 to the vacuole is restored in snc, snc tlg, and temperature-shifted snc1^ala43cells by CAPP activation. (A) Delivery of FM4-64 to the vacuole is restored in snc tlg cells. SNC1, snc (JG8), snc tlg1 (JG9-TLG1), and snc tlg2 (JG9-TLG2) cells were grown to log phase on galactose-containing medium, before being shifted to glucose-containing medium at 26°C for 36 h, with (+C₂) or without (−C₂) C₂-ceramide. Cells were incubated with FM4-64 (1 μg/ml) and processed for fluorescence and light (Nomarski) microscopy. (B) Delivery of FM4-64 to the vacuole is restored in snc vbm1 (MM1), snc tlg1 vbm1 (JG9-TLG1, VBM1), and snc tlg2 vbm1 (JG9-TLG2, VBM1) cells. Cells were grown on glucose-containing medium for 24 h at 26°C, incubated with FM4-64, and processed for microscopy. (C) Delivery of FM4-64 to the vacuole is restored in ceramide-treated snc1^ala43 cells shifted to restrictive temperatures. snc cells expressing snc1^ala43 (JG8-SNC1A43L) were grown in glucose-containing medium at 26°C and either maintained at 26°C during labeling or shifted for 0.5 h to 37°C, before labeling with FM4-64 at the restrictive temperature. Some cells were treated with C₂-ceramide (+C₂) during growth at 26°C.