Figure 5
From: OTUD4 enhances TGFβ signalling through regulation of the TGFβ receptor complex
OTUD4 regulates SMURF2 auto-regulation. (A) HEK293T cells transfected with MYC-SMURF2 and either FLAG-OTUD4 or FLAG-OTUD4 DD. Cells were stimulated where indicated with TGFβ (100 pM) overnight before lysis. After 48 h cells were lysed and whole cell extracts were probed with indicated antibodies. β-Actin is used as the loading control. (B) HEK293T cells transfected with either MYC-SMURF2 or MYC-SMURF2 CA and FLAG-OTUD4. Cells were stimulated where indicated with TGFβ (100 pM) overnight before lysis. After 48 h cells were lysed and whole cell extracts were probed with indicated antibodies. β-Actin is used as the loading control. (C) HEK293T cells transfected with either MYC-SMURF2 or MYC-SMURF2 29/30 and FLAG-OTUD4. Cells were stimulated with TGFβ (100 pM) where indicated overnight before lysis. After 48 h cells were lysed and blotted precipitates were probed with indicated antibodies. β-Actin is used as the loading control. (D) Spearman’s analysis between OTUD4 mRNA expression and Generic EMT score across a TCGA pan-cancer dataset (n = 12,290). Full-length blots for (A,B,C) are shown in Supplementary Information.
