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. 2019 Nov 11;219(1):e201807189. doi: 10.1083/jcb.201807189

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© 2019 Pandey et al.

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Figure S1. — Topo II inhibitor treatment does not affect SUMOylation machinery. (A) A schematic representation of FRET-based in vitro SUMOylation assay. YPet-tagged Topo IIα CTD was used as a substrate, along with CyPet-tagged SUMO2G and SUMO2 GG isoforms in the reaction. (B) In vitro SUMOylation was performed with different PIASy concentrations and at different time points. a.u., arbitrary unit. (C) In vitro SUMOylation was performed for 30 min with different ICRF-193 and merbarone concentrations. The emission spectrum was recorded at 530 nm from three different experiments (n = 3), and error bars represent standard deviation. Paired t test was performed comparing the reaction with inhibitors and without inhibitors in C. None of the inhibitor-containing conditions showed statistically significant differences from the conditions without inhibitors.