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. 2010 Feb 22;30(9):2280–2292. doi: 10.1128/MCB.01392-09

TABLE 2.

Constructs made for the mammalian two-hybrid assaysa

Procedure Figure Human, mouse pGAL4.3 pACT (protein X) pBIND (protein Y) pcDNA3 (vector) Interaction
A 7C Human, mouse mCherry Empty CEP
Human, mouse mCherry ALIX CEP ++
Human, mouse mCherry TSG CEP +
Human, mouse mCherry TX CEP +++
7D Human, mouse mCherry Empty CEP
Human, mouse mCherry TX CEP +++
Human, mouse mCherry Empty ALIX
Human, mouse mCherry TX ALIX
Human, mouse mCherry Empty TSG +, −
Human, mouse mCherry TX TSG +, −
7E Human, mouse mCherry Empty TX
Human, mouse mCherry CEP TX +++
Human, mouse mCherry Empty TX-AAAX3A
Human, mouse mCherry CEP TX-AAAX3A
7F Mouse mCherry Empty CEP
Mouse mCherry TX CEP +++
Mouse mCherry TX-APPX3Y CEP +++
Mouse mCherry TX-GAPX3Y CEP
Mouse mCherry TX-GPAX3Y CEP ++
Mouse mCherry TX-GPPX3A CEP
Mouse mCherry TX-AAAX3A CEP
B 8A and 9, left Mouse, human mCherry Empty CEP NA
Mouse, human mCherry ALIX CEP Empty ++
Mouse, human mCherry ALIX CEP TX +
Mouse, human mCherry ALIX CEP TX-AAAX3A ++
8B and 9, center Mouse mCherry Empty CEP NA
Mouse mCherry TSG CEP Empty +
Mouse mCherry TSG CEP TX
Mouse mCherry TSG CEP TX-AAAX3A +
8C and 9, right Mouse mCherry Empty CEP NA
Mouse mCherry TX CEP Empty +++
Mouse mCherry TX CEP ALIX +++
Mouse mCherry TX CEP TSG +++
a

The table shows the species from which each protein is derived. In most cases the experiments were performed using both human and mouse TX, ALIX, TSG, and CEP. Boldfacing indicates experiments that are presented in Fig. 7C to F, Fig. 8A to C, and Fig. 9. “Empty” refers to the original pACT and pcDNA vector without insert. For procedure A, the two vectors were cotransfected with the pGL4-mCherry vector. For procedure B, the three vectors were cotransfected with the pGL4-mCherry vector. Under the “pcDNA3 (vector)” column, no additional vector is indicated by “NA.” A pBIND-YFP vector with CEP was used for mammalian two-hybrid analyses instead of a pBIND vector for Fig. 9. The terms “+” and “−” indicate positive and negative interactions, and the number of “+” symbols indicates the strength of the interaction.

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