Fig. 3

Blocking antisense strand-loaded RISC activity mitigates hepatotoxicity. a Study design depicting prevention and treatment of rat toxicity by GalNAc-siRNAs using REVERSIR^TM. b Liver exposures for GalNAc-siRNAs in rat prevention (siRNA-1 and siRNA-4) or treatment (siRNA-5) toxicity studies as assessed by stem-loop RT-qPCR for the antisense strand (AS) at necropsy (nx). c Liver RISC loading with or without REVERSIR^TM treatment as assessed by stem-loop RT-qPCR for the antisense strand at necropsy. d Serum glutamate dehydrogenase (GLDH) levels measured at necropsy. Differences between group means were evaluated for statistical significance using one way ANOVA with post hoc corrections (for multiple siRNAs) in GraphPad Prism 7. ns. not significant; *p < 0.05; ****p < 0.0001. Error bars represent standard deviation of the mean. e H&E staining of liver sections collected at necropsy. Known toxic siRNAs administered alone or with a scrambled, control (Ctr) REVERSIR^TM were associated with hepatocellular degeneration (bracket), single cell necrosis (*), increased sinusoidal cells consistent with Kupffer cell hyperplasia and/or infiltrating leukocytes (#), increased mitoses (^), bile duct hyperplasia with fibrosis (+), and hepatocellular vacuolation (arrow). Co-administration of a complementary REVERSIR^TM decreased the severity of these findings and often limited their distribution. All microscopic liver findings are tabulated in Supplementary Table 5. N = 3 males (6–8 weeks old) per group; qw, weekly dosing; q2d, every other day dosing