Yeast two-hybrid analysis of the interactions between different NS1 proteins and PSD-95. (A) AH109 yeasts were transformed with pGBK-PSD-95 plasmid along with pGAD-NS11, pGAD-NS51 or empty vector pGAD-T7 and plated on SD/-Leu/-Trp (DDO) as well as SD/-Ade/-His/-Leu/-Trp (QDO) agar plates and incubated for 3 days at 30°C. The growth status of AH109 was observed at 72 h. (B) Single AH109 yeast colonies grown on the above DDO plates were picked and transferred to QDO/X-α-gal agar plates and incubated at 30°C for 2 days. The representative colonies were shown. (C) Different NS1-encoding plasmids were transformed into AH109 yeast together with PGBK-PSD-95 plasmid for α-galactosidase activities assays and grown in DDO liquid culture with PNP-α-gal as substrate. Data were from 3 independent experiments. pGADT7-T and pGBKT7-p53 plasmids and pGADT7-T and pGBKT7-Lam plasmids were used as positive and negative controls, respectively.