Figure 2.

Binding of GFP ssODNs to MBD4. A 45-nt oligonucleotide radiolabeled probe corresponding to the region of the pGFPmut vector targeted for repair was incubated with or without ssODNs and with and without recombinant MBD4 at 37°C for 30 min and analyzed by EMSA. The position of the ssODN/probe complexes is indicated by the thin arrow. The thick arrow above shows the position of the supershift achieved by the binding of MBD4 with the ssODN/probe complex. MBD4 was unable to form a complex with the target sequence in the absence of an ssODN bearing a mismatch. No binding of MBD4 to m⁵CpG [as assessed using a radioactively labeled m⁵CpG (m⁵CpG*)] was detected in the absence of the oligonucleotide sequence targeted for repair.