Figure 1.

SUMO1 modifies HOXA10 on its evolutionarily conserved lysine 164 residue. (a) HEK293T cells were co-transfected with Myc-HOXA10 and Flag-SUMO1. After 48 h, the cell lysates were affinity-purified using anti-Flag-agarose and anti-Myc-agarose, respectively, and then blotted with anti-Myc and anti-Flag antibodies. An increased molecular weight of ~30 kDA of the detected HOXA10 was observed when HOXA10 and SUMO1 were coexpressed. (b) Extracts from Ishikawa cells transfected with Flag-SUMO1 or an empty vector were incubated with the anti-HOXA10 antibody, and then blotted with anti-HOXA10 and anti-SUMO1 antibodies. Goat IgG was used as a negative control for the co-IP procedure. (c) HEK293T cells were co-transfected with the indicated plasmid. After 48 h, the cell lysates were subjected to anti-flag-agarose precipitation and then immunoblotted with the anti-Myc antibody. (d) HEK293T cells were co-transfected with the indicated plasmids. After 48 h, the cell lysates were directly subjected to western blot.