Figure 1.

Identification of A_2AR^284–410 interacting proteins by co-IP and PD. (A) cMyc-A_2AR^284–410 expression and co-IP from RAW 264.7 cells were detected using an anti-cMyc antibody. Lysates from mock (M) and cMyc-A_2AR^284–410 transfected (S) cells. Non-binding supernatants of precleared (PC) and immunoprecipitated (IP) complexes. Eluted co-IP complex (IC). (B) Recombinant GST and GST-A_2AR^284–410 proteins were expressed in E. coli BLR strain after induction with isopropyl-β-D-thiogalactopyranoside (IPTG) and were affinity purified from the supernatant (SN) fraction of the cell lysates. Recombinant proteins were eluted using 50 mM glutathione and were stained by Coomassie Brillant Blue G250 after separation by SDS-PAGE. (C) Samples prepared for mass spectrometry analyses of isotype control (Ctr-IP) and cMyc antibody specific (S-IP) complexes.