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. 2020 Mar 9;217(5):e20190888. doi: 10.1084/jem.20190888

Figure S5.

Figure S5.

Trib1 interaction with MALT1 requires the Trib1 N terminus between amino acids 82 and 89. (A) Western blot of lysates from sorted WT or Trib1 cKO CD4 T cells ± stimulation with 1 µg/ml anti-CD3/CD28 for 24 h. Neutrophil whole-cell lysates (WCL) were used as a positive control for C/EBPα expression. (B) Schematic of full-length Trib1 (top) and Trib1-ΔCOP1 (bottom). (C) HEK293T cells were transfected with FLAG-tagged empty-vector control, full-length Trib1, or Trib1-ΔCOP1. FLAG-tagged proteins were immunoprecipitated using anti-FLAG agarose beads, and MALT1 binding was assessed by Western blot. (D) Schematic of full-length Trib1 (top), Trib1-Δ1-82 (middle), and Trib1-Δ1-89 (bottom). (E) HEK293T cells were transfected with FLAG-tagged empty-vector control, full-length Trib1, Trib1-Δ1-82, or Trib1-Δ1-89. FLAG-tagged proteins were immunoprecipitated using anti-FLAG agarose beads, and MALT1 binding was assessed by Western blot. Each row is from the same blot and exposure (white lines: cut unrelated lanes). (F) 32D cells (myeloid cell line expressing C/EBPα) were retrovirally transduced with MigR1 empty vector control, full-length Trib1, or Trib1-Δ1-89. Transduced cells were sorted by GFP, and C/EBPα degradation was assessed by Western blot. WT: CD4-cre+Trib1+/+; KO: CD4-cre+Trib1F/F.