Galactose

For the Y1H assay, the AD fusion construct (pB42AD) was co-transformed into yeast (EGY48) with the pLacZ2μ construct containing a specific promoter, and then were selected on SD/-Trp-Ura agar plates for 72–96 h at 30°C. Subsequently, transformants of yeast were carried out using the selective medium containing raffinose, galactose, and X-gal (Amresco, Solon, OH, United States). For the Y2H assay, the AD-fusion (pGADT7) was co-transformed into yeast (AH109) with the BD-fusion (pGBKT7). Yeast (Y2H) transformants were then selected on selective medium (SD-LWHA) with or without 10 mM 3-amino-1,2,4-triazole (3-AT).

The S. cerevisiae strains used in this study are derived from BY4742 (MATahis3Δ1 leu2Δ0 ura3Δ0), obtained from Euroscarf. All yeast strains used in this work are listed in Table 1. YPD medium contained 1% yeast extract (BD, USA), 2% peptone (BD, USA), and 2% dextrose (Amresco, USA). YPG medium contained 1% yeast extract (BD, USA), 2% peptone (BD, USA), and 2% galactose (Amresco, USA). All cultivations were carried out at 30°C.