Figure 4. In vitro translation of PP2A and pull down with a GST construct incorporating the large cytoplasmic loop of the Na⁺,K⁺-ATPase α-subunit.

PP2A A (flag tagged)- and C (HA tagged)- subunit proteins were prepared by in vitro translation and GST pull down was performed with GST alone or GST-Na⁺,K⁺-ATPase large cytoplasmic loop (Na,K-GST). A. GST proteins used for pull down were detected by coomassie brilliant blue (CBB) staining. PP2A C-subunit and A-subunit were detected by Western blot with anti-HA (B) and anti-flag (C) antibodies, respectively. The PP2A C-subunit, but not the A-subunit, specifically co-precipitate with the GST-Na⁺,K⁺-ATPase. Typical results from one of three experiments are shown.