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The data and graphs were then analyzed and generated using GraphPad Prism (v 10.4.1) Differential scanning fluorimetry A total of 9.5 L of the master mix containing FBXO22 FIST WT or variants, SYPRO Orange Dye (Invitrogen), and DSF buffer (20 mM Tris 7.5, 150 mM NaCl, and 1 mM TCEP) were added to a 384-well qPCR plate (Genesee Scientific)