Fig 7. Co-immunoprecipitation assay of adiponectin interaction with p65.

(A) A549 cells treated with curcumin, silenced, or transfected with adiponectin were lysed and adiponectin was co-immunoprecipitated using adiponectin antibody. Immunoblotting with the indicated antibodies confirmed the co-precipitation of adiponectin monomer, dimer, and multimer. (B) Co-immunoprecipitation was performed using anti-adiponectin and anti-p65 antibodies. The p65 elute was separated on SDS-PAGE and immunoblotted with the corresponding antibodies, which showed substantial association of adiponectin with the p65 component. (C) Activation of NF-κB by overexpression or silencing of adiponectin was determined by EMSA. Adiponectin increased the nuclear translocation of p65/p50 and NF-κB activation, but silencing adiponectin expression had the opposite effect.