BiFC analysis of mNT-NAF-1 interaction.
A. Representative images of: (1) Positive control for NAF-1 homodimer interaction using co-expression of NAF-1-YFPc and NAF-1-YFPn with ER tracker localization. (2) Positive control for mNT homodimer interaction using co-expression of mNT-YFPc and mNT-YFPn with mitochondrial (Mito) tracker localization. (3) mNT-NAF-1 interaction following co-expression of NAF-1-YFPc and mNT-YFPn with ER tracker localization. (4) mNT-NAF-1 interaction following co-expression of NAF-1-YFPc and mNT-YFPn with Mito tracker localization. (5) Negative control for mNT-NAF-1 interaction using co-expression of mNT-YFPn and soluble sNAF-1-YFPc with mitochondrial (Mito) tracker localization. B. The different split-YFP/mNT/NAF-1/sNAF-1 vectors used for the in vivo analysis of mNT-NAF-1 interaction shown in A. C. A comparison between the BiFC signal obtained with the mNT-NAF-1 interaction (co-transfection with NAF-1-YFPc and mNT-YFPn; mNT-NAF-1) and the BiFC signal obtained with the mNT-sNAF-1 interaction (co-transfection with sNAF-1-YFPc and mNT-YFPn; mNT-sNAF-1). Vector construction, transfection and imaging are described in Materials and Methods. ***, p≤0.001.