Figure 2

Analysis of the interaction between mCry2 N- and C-termini with mBMAL1 and mPER2 proteins using a mammalian two-hybrid system. ( A) Both mCry2 N- and C-termini cDNA were cloned into pACT, where they are fused with the VP16 domain through their N-termini. The cDNA of mPer2 and mBmal1 were cloned into pBIND and proteins were expressed with the GAL4 domain in HEK 293T cells. (B) HEK 293T cells were transfected with the indicated constructs along with the pGL5luc reporter plasmid. Reporter activity was examined 24 h after transfection. Relative activities were calculated based on mock samples and values were plotted. Error bars indicate SEM from at least 3 experiments. ** p < 0.005, *** p < 0.0001 as determined by student's t-test compared to the relevant controls.