Figure 5 | Cell Death & Disease

Figure 5

From: Functional genetics-directed identification of novel pharmacological inhibitors of FAS- and TNF-dependent apoptosis that protect mice from acute liver failure

Figure 5

Chemical inhibition of candidate targets SSTR5 and PPP2R5A increases mouse survival and reduces liver damage following FAS agonistic antibody (JO2) administration. Kaplan–Meier survival curves for NIH Swiss mice (n=10 per group) injected i.p. with (a) Bim 23056 (1 mg/kg) 1 h before injection of JO2 (240 μg/kg; P=0.0007; n=10 per group) or (b) cantharidin (0.25 mg/kg; P=0.0007; n=10 per group) 4 h before JO2. P-values are for comparison of survival at 48 h between groups pretreated with inhibitor before JO2 or given JO2 alone using Fisher’s exact test. (c) NIH Swiss mice were pretreated with PBS (control, left), cantharidin (0.25 mg/kg, middle) or Bim 23056 (1 mg/kg, right) for 1 h, 1 or 4 h, respectively, before injection of FAS agonistic JO2 antibody (240 μg/kg). Five hours later, liver tissue was collected and processed for H&E staining (top row) and detection of apoptotic cells by TUNEL assay (middle) and cells with activated caspase-3 by immunostaining with antibodies against cleaved Casp-3 (Asp175) (bottom). Representative photographs from groups of three mice are shown. (d – f) Chemical inhibition of candidate targets SSTR5 and PPP2R5A in vivo leads to suppression of FAS-mediated caspase activation in the liver and release of ALT from the liver into the bloodstream. (d and e) Bim 23056 (1 mg/kg) and cantharidin (0.06 mg/kg) were injected i.p. into NIH Swiss mice 1 h before i.p. injection of JO2 (240 mg/kg). Cytosolic liver extracts were prepared 3 h later and used for measurement of caspase 8 (d) and caspase-3/7 (e) activity with luminogenic LETD- or DEVD-containing substrates. (f) ALT activity was determined in serum samples collected from NIH Swiss mice pretreated with cantharidin (0.125 mg/kg) or Bim 23056 (2 mg/kg) 1 h before i.v. injection of ConA (15 mg/kg) or treated with ConA alone. Serum was collected 20 h after ConA injection. For d–f, ‘control’ samples were from animals that received no treatments and the values shown are the means of at least three independent experiments. Error bars indicate standard deviations. P-values were calculated using a moderated student’s t-test

Back to article page