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. 2012 Jan 30;158(4):1628–1642. doi: 10.1104/pp.111.191965

Figure 2.

Figure 2.

Ubiquitin ligase activity tests of LOG2 and LULs. A, In vitro ubiquitination assay of GST-LOG2 and GST-LUL1 to -4 visualized with an anti-ubiquitin antibody. −E2 and −E3, Omission of ubiquitin-conjugating enzyme (E2) and the indicated ubiquitin ligase (E3) from the assay, respectively; T, all ubiquitin pathway components were present in the assay. B, In vitro ubiquitination assay of flag-cGDU1 by GST-LOG2, GST-LUL1, or no added E3, visualized with anti-flag antibody. C, In vitro ubiquitination assay of flag-cGU1 by truncated and mutant forms of GST-LOG2. ΔN, LOG2 truncation lacking amino acids N terminal to DAR2; M, LOG2CC354/347AA (RING-dead); I-A, LOG2I321A (RING-E2 interaction impaired). D, In vitro ubiquitination assay of either flag-cGDU1 or flag-cGDU1G100R by LOG2-V5. The arrowhead indicates nonubiquitinated flag-cGDU1; the pound symbol indicates ubiquitinated flag-cGDU1. Numbers on the left indicate molecular mass in kD. WT, Wild type.