Fig. 2.

Validation of multi-SUMO–binding interactions. (A) GST pulldown analysis of PTRF, BLM, and ZMYM2 binding to the indicated GST-fusion proteins. Epitope-tagged PTRF, BLM, or ZMYM2 were overexpressed in HEK293T cells, and total cell lysates were used for the pulldown. Proteins were detected by IB using anti-FLAG (PTRF) or anti-Myc (BLM and ZMYM2) antibodies. Arrows indicate the positions of bands corresponding to the full-length proteins. Bait proteins were stained with Ponceau (Bottom). (B) Interaction of ZBTB33 or ZBTB4 with COMP-SUMO3 was analyzed by GST-pulldown. Total cell extracts of transfected HEK293T cells were used for the pulldown. Input and precipitated ZBTB33 and ZBTB4 were detected using an anti-FLAG antibody. Proteins used as bait were stained with Ponceau (Bottom).