Cell viability assay
1 × 104 primary mouse or human hepatocytes were
cultured in 96-well plate and transfected with 0.025-0.1 μg of LNP-IL-21
or LNP, or left untreated. Cell viability was assessed at day 1-4 post
transfection with using a Cell Counting Kit-8 kit (Dojindo, Japan)
according to the manufacturer’s instructions.
RNA
sequencing analysis
BALB/c mouse was injected with LNP or 2 × 1011 genome
equivalents (geq) of AAV in 200 μl PBS via tail vein, or left untreated.
At 3 days post injection, RNA extracted form liver tissues was subjected
to library construction and deep sequencing on Illumina NovaSeq 6000 by
Novogene Biotechnology Co. Ltd. (Shanghai, China). FPKM, fragments per
kilobase of transcript per million mapped reads, was used to estimate
gene expression levels. Differential expression analysis of two groups
was performed using the DESeq2 R package (1.20.0). Differentially
expressed genes (DEGs) with a fold change ≥ 2 and a false discovery rate
(FDR) < 0.05 were identified and presented in Table
S1 and S2 .