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. 2018 Dec 26;40:406–421. doi: 10.1016/j.ebiom.2018.12.031

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© 2018 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 2 — Liver Kinase B1 (LKB1) is SUMOylated by SUMO-2 in human hepatoma cells. a. Ni²⁺-NTA agarose bead pulldown in Huh-7 human hepatoma cells after transfection with His-SUMO-1, 2, or 3, with the pcDNA3-FLAG-LKB1 Wild type plasmid (LKB1 WT) in the presence and absence of ubiquitin conjugating enzyme 9 (UBC9). b. Ni²⁺-NTA agarose bead pulldown in Huh-7 human hepatoma cells after transfection with His-SUMO-2 with the LKB1 WT plasmid in the presence of the different SUMO E3 ligases PIAS 1, 2α, 2β, and 4. c. Ni²⁺-NTA agarose bead pulldown in Huh-7 human hepatoma cells after transfection with His-SUMO-2 and with the LKB1 WT plasmid in the presence of the different SUMO-specific proteases, SENPs 1–7. d. Ni²⁺-NTA agarose bead pulldown in Huh-7 human hepatoma cells after transfection with His-SUMO-2 and with the LKB1 WT plasmid or the pcDNA3-FLAG-LKB1 Kinase Dead K78I plasmid. e. Immunoprecipitation assay between LKB1 and the STe20-Related ADaptor (STRADα) cofactor after STRADα overexpression in Huh-7 human hepatoma cells. Normalized quantifications relative to inputs are shown below the panel.