Fig. 3

Optimization of low antigenic and hepatocyte-targeted LNP-based siRNA delivery system. a Schematics of the preparation procedures of LNPs. b Different formulations of LNPs (abbreviated as 1.5% OH, 3% OH, 1.5% OMe and 3% OMe) were prepared, and the sizes, polydispersity index (PDI), ζ potential and encapsulation efficiency (EE%) were analyzed. c C57BL/6 mice were intravenously injected with different formulations of LNPs/Cy5-siRNA at the dose of 0.5 mg/kg, 4 h post-injection, the hepatocytes and liver-associated lymphocytes were isolated, and the percentage of Cy5-positive hepatocytes, T cells, B cells and Kupffer cells (KCs) were determined by flow cytometry analysis. Data were shown as means ± SDs (n = 4). d C57BL/6 mice were intravenously injected with different formulations of LNPs/siApoB at the doses of 0.5 mg/kg and 0.05 mg/kg, mice were sacrificed at 72 h post-injection. Relative ApoB mRNA expression levels in the liver of mice (compared with β-actin mRNA levels) were determined at 72 h post-injection, and (e) the sera total cholesterol (T-CHO) levels were detected via commercial kits. Data were analyzed using two-way ANOVA with Sidak multiple comparison correction, and shown as means ± SDs (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001