Fig. 2
From: The laforin/malin E3-ubiquitin ligase complex ubiquitinates pyruvate kinase M1/M2
The laforin/malin complex polyubiquitinates PKM isoforms attaching K63-linked ubiquitins. a and b HEK293 cells expressing the indicated plasmids were analyzed for in vivo ubiquitination as indicated in Methods. Proteins bound to the metal affinity column (Bound) and those present in the crude extracts (CE, 25 μg) were analyzed by western blotting using the indicated antibodies. The position of polyubiquitinated proteins is indicated. The fuzzy bands in the CE are probably due to the presence of 6 M guanidinium hydrochoride in the extracts