Figure 2.

TAK1 is activated by wild-type H11/HspB8 but not its non-killing mutants. (a) Pull-down assays of untreated and Dox-treated (5 μg/ml; 3 days) stably transfected A375 and A2058 cells with H11/HspB8 or TAK1 antibodies or preimmune (Pre) IgG. (b) Immunocomplex PK assay of cell extracts as in (a) using TAK1 antibody and the TAK1 substrate MKK6 (PK). Kinase precipitates were blotted with TAK1 antibody (IB). (c) TAK1 was not activated in reciprocal pull-down assay as in (a) and immunocomplex PK assay as in (b) but using protein extracts from MeWo and TAG51 cells that respectively express the non-killing H11/HspB8 mutants, P173H and W51C