SI Guide
Supplementary Figure 1 MiR-122 is quantitatively recovered from a duplex formed with antagomir-122.
Equal amounts of miR-122 and antagomir-122 were annealed in vitro and added to a Trizol/liver homogenate. RNA was extracted and samples were separated on a polyacrylamide gel in the absence of formamide. The data demonstrate that miR-122 can be quantitatively recovered from a miR-122/antagomir-122 duplex using the Trizol protocol with ethanol precipitation. Suppl.Fig. 1.pdf Adobe document 68 KB
Supplementary Figure 2 Annealing antagomir-122 to liver RNA in vitro does not alter miR-122 detection in formamide-containing gels.
Total liver RNA was incubated in the presence of different amounts of antagomir-122. Samples were subjected to Trizol extraction and separated in polyacrylamide gels in the presence or absence of 20% formamide. MiR-122 was detected by Northern blotting. Suppl.Fig. 2.pdf Adobe document 128 KB
Supplementary Figure 3 Sustained silencing of miR-122 after injection of antagomir-122 into mice. a, Dose response relationship of total amount of antagomir-122 injected and silencing activity in liver. b, Time-course study demonstrating that the silencing effect of antagomir-122 is long-lasting. Suppl.Fig. 3.pdf Adobe document 220 KB
Supplementary Figure 4 Antagomirs specifically target miRNAs that derive from the same primary transcript.
Northern blots of total RNA isolated from liver and kidney after injection of antagomir- 192 or antagomir-194. The results demonstrate specific inhibition of miRNAs derived from polycistronic precursors by antagomirs. Suppl.Fig. 4.pdf Adobe document 236 KB
1 Supplementary Table 1 Affymetrix gene expression analysis of genes up-regulated ≥ 1.4- fold in livers of mice treated with antagomir-122 compared to control mm-antagomir-122. Suppl.Table 1.xls Microsoft Excel 124 KB
Supplementary Table 2 Affymetrix gene expression analysis of genes down-regulated ≥ 1.4- fold in livers of mice treated with antagomir-122 compared to control mm-antagomir-122. Probe sets with "absent (A)" to "absent" changes were excluded. Suppl.Table 2.xls Microsoft Excel 100 KB
Key genes
Hfe2 hemochromatosis type 2 (juvenile) Tmed3 transmembrane emp24 protein transport domain containing 3 Slc35a4 solute carrier family 35, member A4 AldoA aldolase A, fructose-bisphosphate Ndrg3 N-myc downstream regulated gene 3 Cs citrate synthase Iqgap1 IQ motif containing GTPase activating protein 1 Hmgcr 3-hydroxy-3-methylglutaryl-Coenzyme A reductase Dhcr7 7-dehydrocholesterol reductase Acas2 acetyl-Coenzyme A synthetase 2 (ADP forming) Mvk mevalonate kinase Hmgcs1 3-hydroxy-3-methylglutaryl-coenzyme A synthase 1 Fdps farnesyl diphosphate synthetase Sqle squalene epoxidase Fdft1 farnesyl-diphosphate farnesyltransferase 1
2