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. 2020 Mar 30;219(5):e201907079. doi: 10.1083/jcb.201907079

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© 2020 Sakakibara et al.

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Figure 7. — Formation of the afadin–αE-catenin–β-catenin complex in solution in vitro. (A) Sucrose density gradient centrifugation analysis. MIII-C or β/αE-cat-CA was incubated with CC-afadin and subjected to a sucrose density gradient centrifugation analysis. Each fraction was separated by SDS-PAGE and analyzed by CBB staining. The single asterisk indicates the position of the 1:1 complex of CC-afadin and MIII-C. The double asterisk indicates the position of the 2:2 complex. The dagger indicates the position of the 1:1 complex of CC-afadin and β/αE-cat-CA. The double dagger indicates that of the 2:2 complex. The results shown are representative of three independent experiments. (B) Gel filtration analysis. The fragment of MIII-C was incubated with the fragment of CC-afadin and subjected to a gel filtration analysis. Each fraction was separated by SDS-PAGE and analyzed by CBB staining. Fr. No., fraction number; arrows, CC-afadin; arrowheads, MIII-C; and double arrowheads, β/αE-cat-CA.