Supplementary Figure 3: Analysis of roquin cleavage by MALT1.
(a) CD4+ T cells from Rc3h1fl/flRc3h2fl/flCAG-CARstop-flCd4-Cre mice were infected with empty adenovirus or reconstituted with roquin-1 wild-type, roquin-1(R510G,R579G) or roquin-1(aa1–510) using IRES–GFP co-expressing adenoviruses. Equally infected cells were analyzed as documented by the histogram derived from pregated GFP+ cells. (b) Detection of roquin C-terminal and N-terminal cleavage fragments. CD4+ T cells were isolated from wild-type or Rc3h1fl/flCd4-Cre mice and were preincubated for 4 h with leupeptin (200 μM), MG132 (25 μM) or with DMSO as a control and then stimulated for 60 min with PMA and ionomycin (PMA + iono) or left untreated (–). Cell extracts were prepared and analyzed for roquin protein expression in immunoblots by either using the polyclonal roquin-1 antibody (upper panel) or the monoclonal roquin antibody (lower panel). Data are representatives of two (b) or three (a) independent experiments.
