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Cells were serially diluted and were plated onto rich or minimal medium containing the desired concentration of gliotoxin, using a multi-pronged replicator

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Ubiquitination assay Ubiquitination reactions were performed in a 20 L volume containing the following components: 100 nM human E1 ligase UBE1 (R&D Systems), 500 nM E2 ligase UBC-18, 500 nM GST-tagged WWP-1 or HECD-1 (ProteoGenix), 500 nM His 6 -T7-tagged NHR-49, and 50 M ubiquitin (R&D Systems)