Figure 2. Co-precipitation of cytoplasmic fragments and AMPK.

COS-7 cells transfected with AMPK (GST–α1β1γ1) and GFP–NKCC2^N−term or GFP–NKCC2^C−Term showed reciprocal association with each other. Transfected cell lysates were either immunoprecipitated using an anti-GFP antibody (A and C) or affinity-purified using glutathione-Sepharose (B and D). The anti-GFP immunoprecipitation was probed for AMPK (A) and the glutathione-Sepharose pulldown blotted for GFP (D). The blots were stripped and re-probed to measure the relative expression levels of (B) GST-AMPK and (C) GFP–NKCC. i.p., immunoprecipitation.