Figure 7.

LAS1L is modified by SUMO in a SENP3-controlled manner. (A) LAS1L was generated by in vitro transcription/translation and incubated with recombinant E1 and E2 enzymes and SUMO1 or SUMO2, respectively, in the presence of ATP. In the control reaction (lane 1) SUMO was not added. (B) HeLa cells were transfected with siRNAs and plasmids as indicated. Expression of the respective proteins was verified by western blotting. Detection of β-tubulin served as loading control. (C) HeLa cells were transfected with siRNAs and plasmids as indicated. His-SUMO2 conjugates were captured on magnetic Ni-NTA beads and subjected to western blotting using anti-LAS1L antibodies. Vertical lines indicate removal of irrelevant neighbouring lanes from the initial gel (see Supplementary Figure 14).