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. 2012 Jun 5;7(6):e38294. doi: 10.1371/journal.pone.0038294

Figure 5. Different types of ISG15 modification of endogenous Ubc13.

Figure 5

HeLa cells were transiently transfected with pCMVb-HA-Ubc13 WT (A) or pCMVb-HA-Ubc13 C87G (B) mutant and other vectors as shown in the figure. 24 h post-transfection the cells were collected and lysed in presence 2-ME. The metal-chelate pull-downs were carried out under denatured conditions with or without 2-ME. Immunoblotting against the S-tag show the levels of ISG15. Equal loading of total protein was verified by anti alpha-tubulin immunoblotting.