Fig. 6

TNFAIP8 promotes ERK activation by interacting with its upstream factor Rac1. a, b The interaction of TNFAIP8 and Rac1 was assessed by co-immunoprecipitation experiments in 293 T cells and AML cells, K562 and HL60. Immunoprecipitates were generated by anti-Flag antibody and TNFAIP8-bound Rac1 was detected by immunoblotting with anti-Rac1 antibody. c The effect of TNFAIP8 on Rac1 activation. The levels of Rac1-GTP were determined by a PAK-GST pull-down assay in K562, HL60 cells transduced with Flag-tagged TNFAIP8 or control vector, and corresponding resistant cell lines K562/A02 and HL60/ADR transduced with shTNFAIP8 or shNC which were all stimulated with ADM for 4 h. d Rac1 inhibition by EHOP-016 (5 μM, 12 h) attenuated the positive effect of TNFAIP8 overexpression on ERK phosphorylation