Extended Data Fig. 6: In vitro assays of TXNL1.
a, SDS-PAGE and Coomassie staining of purified HA-tagged TXNL1 variants (left) and TXNL1-deficient human proteasomes (right), representative of 2 independent purifications. b, Coomassie stain of samples from Fig. 2b, representative of 3 independent experiments. IP, immunoprecipitation. c, Purified HA-TXNL1 variants were immunoprecipitated in the presence of purified proteasomes pre-incubated with 6 mM of the indicated compound for 30 min at room temperature before immunoblotting, representative of 2 independent experiments. d, Proteasomes were pre-incubated with TXNL1 or the PSMD14/Rpn11 inhibitor 1,10-phenanthroline for 30 min at room temperature before the addition of a polyubiquitylated (polyUb) sfGFP protein substrate for 10 min at 37 °C, representative of 2 independent experiments. Note: 1,10-phenanthroline inhibits deubiquitylation and degradation of the polyUb-substrate (lane 3), but an excess of TXNL1 does not (lane 4).
