Skip to main content
Figure 1 | BMC Molecular Biology

Figure 1

From: Identification of two Amino Acids in the C-terminal Domain of Mouse CRY2 Essential for PER2 Interaction

Figure 1

Analysis of the interaction between mouse CRYs and various clock proteins using a mammalian two-hybrid system. (A) Mouse Cry2 cDNA was cloned into pACT, where it is fused with the VP16 domain through its N-terminus. The cDNA of mPer2, mClock, 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 pBIND-mBmal1/pACT-mClock, pBIND-mBmal1/pACT-mCry2 and pACT-mCry2/pBIND-mClock, and pACT-mCry2/pBIND-mPer2 separately along with the pGL5luc reporter plasmid. Top Panel: Expression levels of HA-mCRY2 and mCLOCK analyzed by western blot. Bottom Panel: 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. pACT-AD: pACT carries VP16; PBIND-BD: pBIND carries GAL4.

Back to article page