Quantifying Gene Expression in Zebrafish Liver

The quantitative polymerase chain reaction (qPCR) method was used to detect the mRNA expression level of related genes as previously described (18 (link)). Briefly, the livers of zebrafish infected with A. hydrophila were extracted after intraperitoneal injection of bacterial strains for 3 days. Total RNA from zebrafish liver was extracted by the Trizol-chloroform (TaKaRa Inc., Japan) method. The total RNA was then reverse transcribed into cDNA following the instructions in the Prime Script RT Kit (TransGen Biotech Co., Ltd., Beijing, China). Finally, a real-time PCR detection system (Bio-Rad Inc., CA, USA) was used to detect the expression of related genes by real-time qPCR, and 16S rRNA was used as the internal reference gene. The primers used are shown in Supplementary Table S1.