Enrichment and analysis of the NRON complex with size exclusion chromatography.

(A) Methodology for size exclusion chromatography. Extracted and dialyzed proteins were used to enrich the NRON complex through Superdex 200 Increase 10/300 GL size exclusion chromatography column. Created with BioRender.com (B) Molecular weight chromatogram for Superdex 200 Increase 10/300 GL size exclusion chromatography column. GE Healthcare Gel Filtration Calibration HMW Kit is used to prepare the chromatogram. (C) Fraction analysis with western blot to determine the fraction containing the NRON complex components. IQGAP1, CRY1, CSNK1ε, and GSK3β are detected in the collected fractions by Western blot. (D) Stain-free loading control for the blot in Fig 3A. (E) Cytoplasmic and nuclear fractionation verification with Western blot. Tubulin and Histone-H3 are markers of cytoplasmic and nuclear fractions, respectively. (F) CRY1 in the protein lysates of cytoplasmic and nuclear fractions before the SEC experiment. (G) PSMD12 and PSMD4 levels at cytoplasmic and nuclear fractions within the NRON complex.
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