GCGR ASOs decrease GCGR mRNA and lower plasma glucose in ob/ob mice. Mouse (A) or rat (B) primary hepatocytes were treated with the indicated ASO concentration for 4 hours as described in Methods. Following oligonucleotide treatment, cells were cultured for an additional 16–18 hours. Total RNA was extracted, and GCGR mRNA expression was assessed by RT-PCR. GCGR expression was normalized to total RNA in the same samples using Ribogreen. Data are expressed as percent of saline-treated controls. (C) Nonfasted plasma glucose in 8-week-old male ob/ob mice treated twice per week (every 3.5 days) by subcutaneous injection with saline (filled squares), GCGR ASO 148359 (open triangles), GCGR ASO 180475 (open circles), or control ASO 141923 (filled inverted triangles) for 4 weeks. All ASOs were administered at 25 mg/kg. Data are the mean values ± SEM of eight mice per treatment group. In overall comparisons, glucose lowering in animals treated with GCGR ASO 148359 and GCGR ASO 180475 was significantly different compared individually with saline- and control ASO–treated animals (P < 0.05 adjusted using Tukey's t test). (D) Liver GCGR mRNA reduction in ob/ob mice. GCGR mRNA was measured by real-time quantitative RT-PCR from livers of ob/ob mice treated for 4 weeks [i.e., end of the treatment period described in (C)] with saline (black bar), GCGR ASO 148359 (light gray bar), GCGR ASO 180475 (white bar), and control ASO 141923 (dark gray bar). GCGR mRNA was normalized to total RNA in the same samples using Ribogreen. Data are the mean values ± SEM of four mice per treatment group (P < 0.05 using Student’s t test).