Ltc1 StART-like domain is essential for retrograde sterol transfer. (A) Ltc1-sfGFP and truncated alleles lacking StART-like or TM domain expressed under control of the weak ppom1 promoter or strong pact1 promoter in ltc1Δ cells, treated or not with CK-666 for 1 h. (B) Filipin remains at the PM of ltc1Δ cells treated with CK-666 for 1 h. (C) WT but not ltc1Δ cells become resistant to AmB during CK-666 treatment. WT data and methodology as in Fig 2 D. (D) Serial dilutions of WT and indicated mutants on EMM-ALU media containing AmB. (E) Serial dilutions of ltc1Δ strains expressing WT Ltc1 or truncated alleles under the control of ppom1 and pact1 on plates containing 0.2 µg/ml AmB. (F) Serial dilutions of ltc1Δ mutant expressing S. cerevisiae LAM proteins under the control of pact1 on plate containing 0.2 µg/ml AmB. Scale bars, 2 µm.
Figure 6.

Ltc1 StART-like domain is essential for retrograde sterol transfer. (A) Ltc1-sfGFP and truncated alleles lacking StART-like or TM domain expressed under control of the weak ppom1 promoter or strong pact1 promoter in ltc1Δ cells, treated or not with CK-666 for 1 h. (B) Filipin remains at the PM of ltc1Δ cells treated with CK-666 for 1 h. (C) WT but not ltc1Δ cells become resistant to AmB during CK-666 treatment. WT data and methodology as in Fig 2 D. (D) Serial dilutions of WT and indicated mutants on EMM-ALU media containing AmB. (E) Serial dilutions of ltc1Δ strains expressing WT Ltc1 or truncated alleles under the control of ppom1 and pact1 on plates containing 0.2 µg/ml AmB. (F) Serial dilutions of ltc1Δ mutant expressing S. cerevisiae LAM proteins under the control of pact1 on plate containing 0.2 µg/ml AmB. Scale bars, 2 µm.

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