The expression of CD14 gene and several other co-expressed genes were verified using quantitative RT-PCR. We performed quantitative RT-PCR as previously described [27]. Briefly, total RNA was extracted from the hippocampus of 10 BXD strains treated with either ethanol or saline. Three samples were collected from each group for each strain. The total RNA from each individual sample was transcribed into cDNA using a reverse transcription Kit (Invitrogen, Carlsbad,CA) following manufacturer’s instruction. The cDNA was used as the template to amplify the specific products for individual genes using the SYBR Green-based real-time PCR on a LightCycler 4800 real-time PCR instrument (Roche Applied Science; Indianapolis, IN). The relative expression of each gene was normalized to β-actin by using the ΔΔ2Ct method, and data were presented as mean ± SD based on the average of expression levels calculated from all 10 strains by comparing the ethanol group to the saline group. The sequences of the PCR primers are listed in Table 1.
