Fig. 3

Ss-NOBP-1 interacts with Ss-PNO-1. a Ss-NOBP-1 interacts with Ss-PNO-1 in the yeast two-hybrid system. Yeast (Saccharomyces cerevisiae strain Y190) containing pGADT7-Ss-NOBP-1 as bait and pGBKT7-Ss-PNO-1 as prey, or pGADT7-Ss-PNO-1 as bait and pGBKT7-Ss-NOBP-1 as prey, was grown for 48 h on synthetic defined (SD) medium that lacked amino acid (Trp, Leu, His or Ade) and was assayed for LacZ expression for α-galactosidase activity (α-gal). The vectors pGBKT7-Lam and pGADT7-T were used as negative control. pGBKT7-P53 and pGADT7-T were used as positive control. Blue color indicates interaction. b BiFC experiment was performed to check the interaction between Ss-PNO-1 and Ss-NOBP-1 in mammalian cells. Ss-NOBP-1 fused to KN151 and Ss-PNO-1 fused to LC151 were coexpressed in NIH/3T3 cells. pbJun-HA-KN151 and pbFos-Myc-LC151 were coexpressed as a positive control. pHA-KN151 and pMyc-LC151 as a negative control and Blank have no transfection control cells. Punctate red fluorescence (BiFC signal) in the cell indicates interaction. Scale bar = 100 μm