Skip to main content
. 2016 Oct 4;17(11):1624–1640. doi: 10.15252/embr.201642378

Figure 3. Endogenous linear Ub chains are present in Drosophila .

Figure 3

  1. Endogenous linear Ub chains enriched in total protein extracts (input, lane 1) from adult w1118 (w −) flies by GST‐Linear‐TUBE (lane 3). The enriched samples were further treated with recombinant DUBs, vOTU (lane 4), or OTULIN (lane 5). Ub chains were visualized by immunoblotting using anti‐linear Ub antibody or anti‐Ub antibody. Loading of GST proteins was analyzed by Ponceau S staining. *: nonspecific band.
  2. Endogenous linear Ub chains in total cell lysate (TCL) of S2 cells expressing human Flag‐OTULIN WT or catalytically inactive C129A mutant. TCLs enriched with GST‐Linear‐TUBE were examined by immunoblotting using anti‐linear Ub antibody. Expression of OTULIN was analyzed by anti‐Flag antibody, and loading of TCL was detected by anti‐tubulin antibody. Input of GST proteins was analyzed by Ponceau S staining. *: nonspecific band.
  3. Mass spectrometry analysis using an identical sample as in panel (B) lane 6. MS/MS spectra of the prototypic linear Ub chain peptide (GGMQIFVK) is shown.