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A cross-linking step [2 mM 3,3-Dithiodipropionic acid di(N-hydroxysuccinimide ester) (DSP, Sigma-Aldrich) for 30 minutes at room temperature followed by quenching with 20 mM Tris HCl for 15 minutes and 3 PBS washes] was performed prior to cell lysis for the SPHKAP-EGFP co-immunoprecipitation experiments to preserve protein-protein interactions despite proteolytic cleavage of the full-length fusion protein during the IP process
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For the vast majority of patients, these sensations are transient and resolve either as they stay on a consistent dose or once the medication is discontinued
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