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BiFC analysis of mNT-NAF-1 interaction.

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posted on 2017-04-20, 17:40 authored by Ola Karmi, Sarah H. Holt, Luhua Song, Sagi Tamir, Yuting Luo, Fang Bai, Ammar Adenwalla, Merav Darash-Yahana, Yang-Sung Sohn, Patricia A. Jennings, Rajeev K. Azad, Jose' N. Onuchic, Faruck Morcos, Rachel Nechushtai, Ron Mittler

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.

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