*4.1. RNA Extraction, RT-PCR, and qPCR*

Placentas and livers were isolated from WT pregnant mice at E17.5. The samples were immersed in TRIzol reagent (Ambion, Austin, TX, USA) and lysed to extract RNA using Qiagen tissue lyser II, as previously described [5]. RNA was then reverse transcribed into cDNA using a Reverse Transcription Master Premix (Elpis Biotech, Daejeon, Korea) for RT-PCR and qPCR. RT-PCR was performed at an annealing temperature of 64 ◦C for Stab1 and 60 ◦C for Stab2 (35 cycles), and 60 ◦C for Gapdh as a loading control (25 cycles). Results of RT-PCR were analyzed using ImageJ (National Institute of Health, Bethesda, MD, USA). qPCR was performed using Applied Biosystems StepOnePlus qPCR system (Life technologies Software v2.3; Life technologies, Carlsbad, CA, USA) under the following conditions: initial denaturation at 95 ◦C for 10 min, 40 cycles of denaturation at 95 ◦C for 15 s and amplification at 60 ◦C for 1 min, and a final cooling step. Results of qPCR were analyzed using the comparative cycle threshold (*CT*) method. Primer sets used for this experiment are as follows: *Stab1*, 5 0 -TGC GAC ATC CAC ACC AAG TT-30 and 50 -TGA ACC ACA TCC TTC CAG CA-30 ; *Stab2*, 50 -AGC TGC TGC CTT TAA TCC TCA-30 and 50 -ACT CCG TCT TGA TGG TTA GAG TA-30 ; and *Gapdh*, 5 0 -GCA TCT CCC TCA CAA TTT CCA-30 and 50 -GTG CAG CGA ACT TTA TTG ATG G-30 .
