Extended Data Figure 8: DUXBL interacts with the TRIM33/TRIM24 complex.

a, b) Western blot analysis of the indicated proteins performed with total protein extracts (input) and DUXBL immunoprecipitates (10% of total IP) obtained from untreated or DOX-treated ESC^DUXBL (a) or untreated or DOX-treated ESC^DUX (b). c) Volcano plot showing the enrichment of proteins obtained from endogenous DUXBL immunoprecipitation followed by mass spectrometry (IP-MS) analysis in two independent WT untreated or DOX-treated ESC^DUX. d) Western blot analyses of the indicated proteins performed with total extracts (input) and DUXBL immunoprecipitates (IP) obtained from untreated and doxycycline treated ESC^DUX. We also included the flowthrough (FT) following the immunoprecipitation to show the specific depletion of DUXBL where corresponds. e) Schematic representation of the different TRIM24 mutants used in our immunoprecipitation assays. f, g) Western blot analysis of the indicated proteins performed with total protein extracts (input) and FLAG pulldowns obtained from DOX-treated ESC containing stably transfected Piggy-bac constructs encoding for HA-tagged DUXBL and/or each of the different FLAG-tagged TRIM24 mutants. h) Schematic representation of the different DUXBL protein forms used in our immunoprecipitation assays. i) Western blot analysis of the indicated proteins performed with total protein extracts (input) and FLAG pulldowns obtained from DOX-treated DUXBL^KO ESC containing stably transfected Piggy-bac constructs encoding for each of the different FLAG-tagged DUXBL protein forms. TRIM24 endogenous levels are shown. Two independent experiments were performed in (a, b, d, f, g and i) but one representative is shown.