Gel mobility shift (GMS) assay with gus-PSTVd-substrates and with the 75-nt R/G site stem-loop structure. (A) GMS assay with gus-PSTVd-R/G75. Increasing amounts of recombinant ADAR2 were added as indicated. A shift to a higher-molecular-weight band was found at 0.17 nM protein. (B) GMS assay with gus-PSTVd; the amount of ADAR2 added is indicated. In lanes marked as “post phenol,” the samples were treated with phenol after protein binding (see Materials and Methods). Lanes marked as “post editing” indicate an editing assay prior to the binding assay. (Lane C) A control with nonedited free RNA. (C) GMS assay with a 75-nt-long R/G site stem-loop structure. Increasing amounts of recombinant ADAR2 were added as indicated.
