(A) Laf1 has a function in retrograde sterol transport. Serial 10-fold dilutions of WT GFP-Lam2 (YFR512-A), laf1∆ GFP-Lam2 (YFR603), dgr2∆ GFP-Lam2 (YFR607), and laf1∆ dgr2∆ GFP-Lam2 (YFR613-A) were spotted on plates lacking (-) or containing AmB at the indicated concentrations. The plates were scanned after 2 days of growth at 30 °C. (B) Laf1 is more abundant than Dgr2. Extracts of cells expressing, as indicated, either Laf1-mKate (YFR635) or Dgr2-mKate (YFR657-A), each from their endogenous chromosomal locus, were resolved by SDS-PAGE and analyzed by immunoblotting, as described in the Materials and Methods Section. (C) Laf1 functions in the same retrograde sterol transport pathway as Lam2 and Lam4. Serial 10-fold dilutions of WT (BY4741), lam2∆ lam4∆ (YFR513), laf1∆ (ymr102c∆), and lam2∆ lam4∆ laf1∆ (YFR734-A) were spotted on plates lacking (-) or containing AmB at the indicated concentrations. The plates were scanned after 2 days of growth at 30 °C.