2.6. Gene expression analysis
Genes including ser-1 , mod-1 , ser-6 , acs-4 ,dop-3 and cpt-1 are directly related with neural signals and lipid metabolism (Lemieux and Ashrafi, 2016; Srinivasan et al., 2008; Watts, 2009). Their expression levels were determined with real-time polymerase chain reaction (RT-PCR) (Li et al., 2020). Briefly, TRIzol reagents were used to extract the total RNA which was further synthesized to cDNA for subsequent RT-PCR with SRBR Premix Ex Taq™ (Takara Bio Inc., Japan) in FTC-3000 (Funglyn Biotech Inc., Canada). The expression levels of chosen genes were quantified by the 2-ΔΔCT method with gpd-2 as the endogenous reference gene. The primers are listed in Table S1 in Supporting Information.