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. 2018 Dec 8;47(3):1070–1081. doi: 10.1093/nar/gky1232

Figure 5.

Figure 5.

Mechanistic analysis of lipid-hsiRNA internalization in liver. (A) Guide strand quantification of Cy3-labeled DHA-hsiRNAs and DCA-hsiRNAs in wild-type (C57BL/6J) and LDLR deficient animals after a single, intravenous injection (n = 3 mice, 10 mg kg−1) using a PNA hybridization-based assay. Data presented as mean ± SD. (B) Hepatocyte internalization of Cy3-labeled DCA-hsiRNA in wild-type and LDLR deficient animals after a single, intravenous injection (n = 3 mice, 10 mg kg−1). Image is representative. Cy3-labeled DCA-hsiRNAs (red), nuclei stained with DAPI (blue). (C) Quantification of fluorescent signal from images acquired in (B). (D) Average retention times of Cy3-labeled DCA-hsiRNAs in mouse serum, 15 minutes after IV injection (n = 2 mice, wild type or LDLR−/−). (E) Average peak integrations from lipoprotein profiles in (D). (F) Hepatocyte internalization of Cy3-labeled blunt and asymmetric siRNAs (unconjugated, DCA-conjugated, or GalNAc-conjugated) after a single, subcutaneous injection (n = 3 mice, 20 mg kg−1), staining as described in (B). (G) Quantification of fluorescent signal from images acquired in (F).