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We first confirmed the active GLP-1RSPHKAP interaction inferred from our MS interactome analysis by measuring increased co-immunoprecipitation of SPHKAP-EGFP (Fig

The eluted proteins were buffer-exchanged to DPBS and concentrated using Amicon Ultra-15 Centrifugation Filter Units (Millipore) with a 30 kDa cutoff by centrifugation at (290400) g at 4 C

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Cell viability was assessed by AlamarBlue turnover (Thermo Fisher Scientific, DAL1025) according to the manufacturers recommendations