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. 2020 May 19;295(27):9244–9262. doi: 10.1074/jbc.RA120.013077

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© 2020 Kim et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 2. — Direct binding of Clstn3 cadherin domains to β-Nrxns. A, diagrams of Clstn3 variants used in pulldown assays presented in B. B, IgNrxn1α^+SS4, IgNrxn1β^+SS4, or IgC (negative control) proteins pulled down FLAG-Clstn3 Full and FLAG-Clstn3 Cad, but not FLAG-Clstn3 ΔCad. Input, 5%. C, IgNrxn1α^+SS4, IgNrxn1β^+SS4, or IgC used for pulldown assays were analyzed by direct comparison of bands revealed by parallel Ponceau S staining. D, diagrams of His-HA–tagged extracellular Clstn3 variants used in direct-binding assays. E and F, purified His-tagged Clstn3 proteins were incubated with purified IgNrxn proteins, as indicated. Precipitates obtained using Talon resin were analyzed by immunoblotting with human IgG or HA antibodies. Input, 10%.