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. 2015 Oct 10;43(18):9039–9050. doi: 10.1093/nar/gkv845

Figure 5.

Figure 5.

Comparison of the impact of mutations in the canonical and non-canonical backside on Rad6 activity. Ubiquitination of histone H2B in HeLa nucleosomes by wild-type Rad6, canonical backside mutant Rad6S25R and non-canonical backside mutant Rad6L56A. The ubiquitination reaction was performed at 30°C at different time points with 2 μM E2 in the reaction buffer containing 0.1 μM E1, 20 μM ubiquitin, 1.25 μM HeLa nucleosomes. H2B and ubiquitin were probed by Western blotting using the anti-H2B, anti-Ub antibodies.