Fig 1: Proteasome assembly and activity.a Levels of Ubn–protein conjugates in WT Col-0, rpn12a-1, rpt2a-2, rpn10-1, rpn12a-236, and 35S:RPN12a leaves. Twenty micrograms of total protein (control = Coomassie blue staining with a focus on Rubisco large subunit) were subjected to SDS-PAGE and immunoblot analysis with anti-ubiquitin antibodies. The signal intensity of ubiquitin-protein conjugates was estimated, and fold-change as compared to WT Col-0 was reported below the immunoblot for each genotype. b Proteasome chymotrypsin-like activity. Native-PAGE followed by immunoblot analyses of the PBA1 subunit (middle) and activity test (right) in 10 µg of total protein extracts from 10-day-old seedlings. For the 20S proteasome activity, native-PAGE membrane was incubated with a Suc-LLVY-AMC fluorophore solution and revealed at 365 nm. Loading control was checked by Coomassie staining (left) (experiments were run in triplicates from independent extracts).