Fig 2: Differential pTyr-Prdx1/ERK interactions in EL-Kras and KC mouse tissue(A) Levels of pERK bound to pTyr-Prdx1 after immunoprecipitation of pTyr-Prdx1 from whole tissue lysate of EL-Kras and KC mouse pancreas. (B) Levels of Prdx1 bound to pERK after immunoprecipitation of pERK from whole tissue lysate of EL-Kras and KC mouse pancreas. (C) Prdx1 expression and oligomerization in whole tissue lysate from the same EL-Kras and KC mice after post-lysis treatment with DTT. (EL-Kras, n=5; KC, n=4.)

Fig 3: Expression of the Txn system is dysregulated during pancreatic neoplasia in vivoKC and EL-Kras mice were stained for Prdx1 (IHC), Txn (IF), and Srxn (IF). Both normal pancreatic tissue and lesions are represented. N=normal tissue and L=lesions in enlarged inset. (n=5/genotype). (Enlarged pictures in Supplementary Figures 4-6.)

Fig 4: Regulation of the Txn system and its effect on ERK and AKT phosphorylation in primary culture of mutant Kras pancreatic cellsPrimary pancreatic cells were derived from EL-Kras and KC mice and treated with the TxnR inhibitor auranofin. (A,B) Regulation of ERK and AKT phosphorylation by auranofin in (A) EL-Kras and (B) KC mice. (C,D) Auranofin induced changes in Prdx1 expression in (C) EL-Kras and (D) KC mice in non-reducing westerns (EL-Kras n=4, KC n=4). (Statistical analysis shown in Supplementary Figures 10 and 11. Supporting Txn and Srxn data shown in Supplementary Figures 11 and 12.)

Fig 5: pERK and Prdx1 interactions and Txn system regulation of pERK and pAKT in pancreatic cellsThe effect of auranofin on Kras effectors and the Txn system was evaluated in vitro in normal and mutant Kras ductal cells (HPDE and HPDE-Kras) and in primary tumor derived (Panc1) and metastatic pancreatic cancer cell lines (AsPC-1). Auranofin induced (A) Prdx1 expression in non-reducing westerns, (B) ERK and AKT phosphorylation, and (C) total protein cysteine oxidation (CSA) in non-reducing westerns. (D) pERK and total ERK protein was immunoprecipitated then probed for Prdx1 binding in reducing westerns (westerns n=7, immunoprecipitations n=3). (Statistical analysis shown in Supplementary Figures 8 and 9.)