Supplementary Materials

This PDF file includes:

  • Fig. S1. Discovery of multiple PKIS compounds as TRIB2-binding compounds.
  • Fig. S2. Chemical structure of preclinical and clinical compounds evaluated in this study.
  • Fig. S3. Validation of DSF assay using TRIB1 and a PKAc counterscreen.
  • Fig. S4. Thermal melting profiles of TRIB2 using DSF.
  • Fig. S5. Analysis of C104Y mutation in TRIB2.
  • Fig. S6. MS-based analysis of the covalent TRIB2:afatinib complex.
  • Fig. S7. MST assay.
  • Fig. S8. Molecular docking analysis.
  • Fig. S9. A new TRIB2 antibody for quantitative analysis of TRIB2 expression levels and stability.
  • Fig. S10. Analysis of TRIB2 dephosphorylation in cell extracts.
  • Fig. S11. TRIB2 binding to afatinib induces destabilization relative to DMSO in a whole-cell thermal shift assay (CETSA).
  • Fig. S12. Comparative protein expression analysis of stable HeLa FLAG-TRIB2 cell lines.
  • Fig. S13. Lack of effect of the noncovalent TRIB1- and TRIB2-destabilizing compound GW804482X on TRIB2 stability in TRIB2-expressing HeLa cells.
  • Table S1. PKIS compound screening data for full-length TRIB2.

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