title: Illumina HiSeq 2000 sequencing; Perturbation of placental endocrine function by endoplasmic reticulum stress leads to maladaptation of maternal hepatic glucose metabolism
design: Perturbation of placental endocrine function by endoplasmic reticulum stress leads to maladaptation of maternal hepatic glucose metabolism
library_construction_protocol: RNA Sequencing was performed using BeWo choriocarcinoma cells.  Cells were treated with the ER-stress induce, thapsigargin (concentration is 100 nM. As thapsigargin was dissolved in DMSO and was at 1 mM stock, therefore, 0.01% DMSO was added as vehicle control.) or vehicle in a paired design for 24 hour in serum-free modified DMEM/F12 medium containing 5.5 mM glucose.  This was repeated using 5 independent replicate cultures (so 10 cultures in total).     thapsigargin (Tg) with/without treatment for BeWo cells Nucleic acid extraction protocol: RNeasy Plus Mini Kit (catalog no. 74134; Qiagen) was used to isolate total RNA following manufacturerâ€™s instruction. Illumina TruSeq Stranded mRNA Library Kit