RNA Extraction and qRT-PCR Analysis

Total RNAs of decidua tissues were extracted using Trizol (Invitrogen) according to manual instructions. Quantitative RT-PCR (qRT-PCR) was performed using the HiScriptII Supermix (Vazyme, Nanjing, China) following the manufacturer’s instructions. Briefly, RNA was quantified using a Nanodrop One instrument (Thermo, MA, USA) and 1 μg was used for reverse transcription using random primers. For qRT-PCR, SYBR Green master mix and primers (final concentration at 200 nM) were used and results were analyzed in CFX Connect PCR detection system (Bio-Rad, CA, USA). Primers were designed according to a previous publication (22 (link)) for CCL8 and GAPDH, or using an open resource (www.ncbi.nlm.nih.gov/tools/primer-blast) for TRDV1, TRDV2, and TRDV3, whose sequences are listed in Supplementary Table S2. Expression levels were normalized to GAPDH and represented as fold change compared to the control (2^−ΔΔCt).