Fig. 5: MYSM1 disrupts TRAF2–TRAF3 complex formation via K63-linked de-ubiquitination. | npj Precision Oncology

Fig. 5: MYSM1 disrupts TRAF2–TRAF3 complex formation via K63-linked de-ubiquitination.

From: MYSM1 promotes lung adenocarcinoma progression via TRAF2/3-mediated activation of MAPK and non-canonical NF-κB pathways

Fig. 5: MYSM1 disrupts TRAF2–TRAF3 complex formation via K63-linked de-ubiquitination.The alternative text for this image may have been generated using AI.

A Venn diagram showed 91 proteins present in the IP group but not in the IgG group in the mass spectrometry analyses, of which MYSM1, TRAF2, and TRAF3 were all present. B Immunoprecipitation was performed in H1299 with high MYSM1 expression to detect MYSM1 interactions with TRAF2 and TRAF3. C Flag-tagged MYSM1 plasmid and HA-tagged TRAF2/TRAF3 plasmid were transfected in 293 T cells, and immunoprecipitation was performed to detect the interactions of MYSM1 with TRAF2 and TRAF3. D The interaction between MYSM1, TRAF2/3 and ubiquitin was assessed by immunoprecipitation after transfection of MYSM1, TRAF2/3, and ubiquitin granules in 293 T cells. E Western blot (WB) analysis of WCL and anti HA pull-down products derived from 293 T cells transfected with Flag-MYSM1, HA-TRAF2 and K-only ubiquitin mutants. Where indicated, 10 μM MG132 was added for 8 h before harvesting the cells. F WB analysis of WCL and anti HA pull-down products derived from 293 T cells transfected with Flag-MYSM1, HA-TRAF2 and the indicated ubiquitin KR (Lys to Arg) mutants. Where indicated, 10 μM MG132 was added for 8 h before harvesting the cells. G WB analysis of WCL and anti HA pull-down products derived from 293 T cells transfected with Flag-MYSM1, HA-TRAF3 and K-only ubiquitin mutants. Where indicated, 10 μM MG132 was added for 8 h before harvesting the cells. H WB analysis of WCL and anti HA pull-down products derived from 293 T cells transfected with Flag-MYSM1, HA-TRAF3 and the indicated ubiquitin KR (Lys to Arg) mutants. Where indicated, 10 μM MG132 was added for 8 h before harvesting the cells. I WB analysis of WCL and anti HA or flag pull-down products derived from 293 T cells transfected with HA-TRAF2, Flag-TRAF3 and Myc-MYSM1 protein constructs. 30 h post-transfection, cells were treated with 10 μM MG132 for 8 h before harvesting. J WB analysis of WCL and anti HA pull-down products derived from control and MYSM1 knockdown 293 T cells transfected with HA-TRAF2 and Flag-TRAF3 protein constructs. 30 h post-transfection, cells were treated with 10 μM MG132 for 8 h before harvesting. K WB analysis of WCL and anti HA pull-down products derived from control and TRAF2 knockdown 293 T cells transfected with HA-TRAF3 and Flag- MYSM1 protein constructs. 30 h post-transfection, cells were treated with 10 μM MG132 for 8 h before harvesting. L WB analysis of WCL and anti HA pull-down products derived from control and TRAF3 knockdown 293 T cells transfected with HA-TRAF2 and Flag-MYSM1 protein constructs. 30 h post-transfection, cells were treated with 10 μM MG132 for 8 h before harvesting. M Truncated forms of MYSM1 were constructed according to its functional domains and co-immunoprecipitation assay of truncated forms of MYSM1-Flag and TRAF2/3-HA in the whole cell lysates of 293 T cells.

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