The effect of C5 deficiency on gene expression in liver tissue of NASH mice
C5 deficiency and wild type (WT) mice were fed WD and injected with low-dose CCl4 for 12 weeks. To explore the putative mechanisms of C5 effects in the progression of NASH, transcriptional profiling of liver tissues from WT and C5 deficient mice was performed.
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C5−/− (B10.D2-Hc0H2dH2-T18c/0SnJ) mice and their haplotype (B10.D2-Hc1H2dH2-T18c/nSnJ) were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). All mice were maintained in a specific pathogen-free facility at Guangxi Medical University. A western diet (WD) and chemically induced murine NASH model as described previously were used. Briefly, mice were fed a WD combined with low weekly dose of intraperitoneal carbon tetrachloride (CCl4) . Male C5−/− and their haplotype, aged 8-12 weeks, were fed WD/CCl4 (WD+CCl4) for 12 weeks. CCl4 were intraperitoneally injected into 8-week-old male mice once a week. Transcriptional profiling of liver tissues from WT and C5 deficient mice was performed. Total RNA was extracted from flash-frozen liver tissues of C5−/− and their haplotype NASH mice with TRIzol reagent (Invitrogen, Carlsbad, CA, USA). The quality of the RNA samples was evaluated with a NanoDrop 2000c spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and an Agilent’s Bioanalyzer. Sequencing libraries were generated by reverse transcription-polymerase chain reaction (RT‑PCR) amplification. The PCR products were sequenced on a HiSeq 2500 sequencing system (RIBOBIO, Guangzhou, China).