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. 2020 Feb 7;7:6. doi: 10.3389/fmolb.2020.00006

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Copyright © 2020 Imperatore, McAninch, Valdez-Sinon, Bassell and Mihailescu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Bi-PSD-95 Q1-Q2, Bi-PSD-95 M2, Bi-Shank1a GQ, and Bi-Shank1a M2 pull down of endogenous FUS and FMRP. The Bi-PSD-95 Q1-Q2, Bi-PSD-95 M2, Bi-Shank1a GQ, and Bi-Shank1a M2 probes were denatured at 95°C in 10 mM cacodylic acid pH 6.5 and 150 mM KCl for 5 min and cooled at room temperature for 15 min. 5 μM of each probe was incubated with E17 mouse brain lysate for 20 min at room temperature and NeutrAvidin agarose (Thermo Scientific, Inc.) pre-blocked with BSA was used to precipitate the probes. After extensive washing, proteins were detected by immunoblot against FUS (top panel) and FMRP (middle panel). GADPH (bottom panel) was used as a control. The pull down experiments were performed in triplicate, the gels shown are representative results from one experiment.