Fig. 4

TNFAIP2 interacting with KEAP1 results in NRF2 stabilization. (a) IP/MS assay of TNFAIP2-interacting proteins. (b) Co-IP assay of the TNFAIP2 and KEAP1 interaction in HNSCC cell lines. (c) Flow cytometry analyses of ROS in TNFAIP2-overexpressing HNSCC cell lines with or without KEAP1 knockdown. (d-e) Cisplatin IC50 evaluations in TNFAIP2-overexpressing FADU (d) and CAL33 (e) cells with or without KEAP1 knockdown. (f) Western blot analysis of NRF2 in TNFAIP2-overexpressing HNSCC cell lines with or without KEAP1 knockdown. (g) Co-IP assay of the amount of NRF2 immunoprecipitated by KEAP1 in TNFAIP2 knockdown HNSCC cell lines treated with MG132 (10 µM, 4 h). (h) Co-IP assay of the amount of NRF2 immunoprecipitated by KEAP1 in TNFAIP2-overexpressing HNSCC cell lines. Data are presented as the mean ± SEM. n.s., not significant; *** P < 0.001. IP/MS, immunoprecipitation coupled with mass spectrometry