Robert Hanczko, David R. Fernandez, Edward Doherty, Yueming Qian, Gyorgy Vas, Brian Niland, Tiffany Telarico, Adinoyi Garba, Sanjay Banerjee, Frank A. Middleton, Donna Barrett, Maureen Barcza, Katalin Banki, Steve K. Landas, Andras Perl
Increased susceptibility of Taldo1–/– mice to liver failure induced by APAP.
(A) Survival of Taldo1+/+ (n = 26), Taldo1+/– (n = 28), and Taldo1–/– littermates (n = 23) injected with 800 mg/kg APAP. Log-rank test showed reduced survival of Taldo1–/– compared with Taldo1+/+ mice (P = 0.027). No significant difference was observed between the Taldo1+/– and Taldo1+/+ groups. (B) H&E-stained liver sections obtained 6 hours after APAP injection. Hemorrhagic necrosis, characterized by hepatocyte vacuolization and extravasation of erythrocytes, was enhanced in Taldo1–/– liver. Original magnification, ×100. (C) Western blot detection of 46- and 54-kDa JNK and their state of phosphorylation (p46 and p54 JNK) in APAP-injected and untreated control mice. Numbers below blots show p-JNK/JNK levels, which were determined relative to actin and normalized to untreated Taldo1+/+ protein lysates, set as 1.0. (D) Assessment of JNK activity by in vitro phosphorylation of GST–c-Jun1–89 fusion protein in APAP-treated liver. TAL, c-Jun, and actin levels were detected by Western blot of liver cell lysates. In vitro phosphorylation of GST–c-Jun was detected by Western blot analysis using anti–phospho–c-JunSer63 antibody. (E) Effect of SP600125 on APAP-induced activation of JNK. Littermate 12-week-old mice were pretreated with SP600125 or DMSO control as described in Methods 1 hour prior to APAP exposure. Numbers below blots show p-JNK/JNK levels 3 hours after APAP treatment; values were determined relative to actin and normalized to untreated Taldo1+/+ protein lysates, set as 1.0.