Fig. 3

TcNST1 cannot complement a S. cerevisiae mutant strain deficient in GDP-Man transport. a S. cerevisiae strain NDY5 was transfected with TcNST1-V5, the S. cerevisiae GDP-Man Vrg4p transporter (positive control) or empty vector (pYEST-DEST 52, negative control). Wild-type S. cerevisiae (JPY25 6c) cells were included in the analysis for comparison. Transfected cells were grown to exponential phase in liquid cultures of SCM-URA, adjusted to an OD of approximately one (lanes 1 and 2) and submitted to ten-fold serial dilution (lanes 3–5). Cells were then inoculated on solid agar plates containing YP plus 2 % galactose and 1.6 μg/ml Congo red. Growth was assessed after 4 days at 30 °C. b Western blot analysis of strain NDY5 transfected with empty vector (lane 1) or TcNST1-V5 (lane 2). TcNST1 expression was detected with a mouse anti-V5 antibody. Expression of histone 3 was used as an internal control