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. Author manuscript; available in PMC: 2010 Oct 14.
Published in final edited form as: J Biol Chem. 2007 Apr 11;282(22):16054–16061. doi: 10.1074/jbc.M611392200

FIGURE 4. Homodimerization of ADAR is independent of dsRNA binding.

FIGURE 4

The ADAR1 (WT and EAA) and ADAR2 (WT and EAA) proteinswere co-expressed in Sf9 cells with its differentially tagged partner. These recombinant proteins were sequentially purified on an anti-FLAGmAb affinity column (left panel) and then on a second TALON affinity column (right panel). Western blotting analysis specific for the first purification using the anti-FLAG M2 mAb indicates the F-tagged protein eluted (left panel) and the anti-His6 mAb reveals the H-tagged protein that is retrieved (right panel), confirming the interaction. Single purified recombinant F-ADAR2 and H-ADAR2 expressed with one tag was included to show the specificity of the two mAbs used forWestern blotting analysis (lanes 1, 2, 7, and 8). These F/H-tagged ADAR1 and ADAR2 oligomeric complexes were alsoidentified with the reciprocal mAb toconfirm the presence of the other monomer subunit partner (not shown). Approximately 10 ng of each purified protein was loaded onto the SDS-polyacrylamide gels.