RT-qPCR was performed using the QuantStudioFlex 12 K instrument (Applied Biosystems). Taqman gene expression assays listed in Table 1 were used according to the manufacturer’s instructions, with cDNA (5 ng) in technical triplicates. The PCR program was as follows: UNG activation (50 °C, 2 min), UNG inactivation (95 °C, 10 min), followed by 40 cycles of denaturation (95 °C, 15 s) and annealing/extension (60 °C, 1 min). The mean Ct values were calculated from technical triplicates. For the calculation of fold change in expression, the ΔΔCt method was used . For samples with Ct values that were greater than 35 was considered a negative, however for the purpose of calculating a fold change, these samples were assigned a Ct value of 35. The mean ΔCt value of the control (untreated) samples was calculated, and individual ΔCt values were calculated for experimental replicates within the LPS treated samples, using the formula: ΔCt = Ct target gene – Ct reference gene. The ΔΔCt value for each LPS treated sample was calculated as the ΔCt LPS sample – mean ΔCt control samples. This was then transformed into a fold change value for each LPS sample using the calculation 2-ΔΔCt.Table 1Taqman gene expression assaysGeneAssay IDDyeActinbβ-actinMm00607939 slFAM-MGBB2mβ-2-microglobulinMm0043762 mlFAM-MGBGapdhGlyceraldehyde-3-phosphate dehydrogenaseMm99999915 glFAM-MGBGusbβ-glucuronidaseMm01197698 mlFAM-MGBHmbsHydroxymethylbilane synthaseMm01143545 mlFAM-MGBHnrnpabHeterogeneous Nuclear Ribonucleoprotein A/BMm1288699 mlFAM-MGBHprtHypoxanthine-guanine phosphoribosyltransferaseMm00446968_mlFAM-MGBMau2MAU2 Sister Chromaid Cohesion FactorMm00512415 mlFAM-MGBPpiaPeptidylprolyl Isomerase AMm02342430 glFAM-MGBRpl13aRibosomal protein L13aMm01612987 glFAM-MGBStx5aSyntaxin 5aMm00502335 mlFAM-MGB