Rhesus specific taqman gene expression primers

Total RNA was extracted from snap-frozen chorioamnion-decidua and amnion biopsies after homogenizing in TRIzol (Invitrogen, Waltham, MA, USA). RNA concentration and quality were measured by NanoDrop spectrophotometer (Thermo Fisher Scientific, Wilmington, Delaware, USA). Reverse transcription of the RNA and quantitative RT-PCR were performed using qScript One-Step RT-qPCR Kit (Quanta BioSciences, Beverly, MA, USA) following the manufacturer’s instructions and with Rhesus-specific TaqMan gene expression primers (Life Technologies). Eukaryotic 18S rRNA (Life Technologies) was endogenous control for normalization of the target RNAs.

Total RNA was extracted from snap-frozen chorioamnion decidua after homogenizing in TRIzol (Invitrogen). RNA concentration and quality were measured by a nanodrop spectrophotometer. Reverse transcription of the RNA was performed using Verso cDNA synthesis kit (Thermo-Scientific), following the manufacturer’s protocol. Quantitative RT-PCR was carried out in a StepOnePlus Real Time PCR system (Life Technologies) following standard cycling conditions. Quantitative RT-PCRs were performed with rhesus-specific TaqMan gene expression primers (Life Technologies). The eukaryotic 18S rRNA (Life Technologies) was endogenous control for normalization of the target RNAs and a sample from a placebo inoculated animal was used as a calibrator. The cytokine mRNA values for ZIKV-infected animals were expressed relative to the average value of the control group. Statistical analysis on cytokine mRNA values were done via t-test, with two-sided p values of <0.05 considered significant.