MOTHER_venous_EC_RNA_sequencing_results.xlsx

The dataset includes venous endothelial cell (EC) transcriptional profiling from 24 postpartum participants, including 17 with severe preeclampsia (mean ± SD age 32.6 ± 2.7 years) and 7 with normotensive pregnancy (32.4 ± 4.0 years), with venous ECs harvested at a mean ± SD 15.4 ± 7.9 and 11.5 ± 9.0 days postpartum, respectively.Venous ECs were harvested from an antecubital vein after IV placement followed by insertion of four J-wires using sterile technique. ECs were collected from the wires by gently washing 10X and centrifuged (400 x g for 5 min at 4 deg C with no brake) in dissociation buffer (phosphate-buffered saline[PBS], supplemented with 0.5% bovine serum albumin [BSA], 2mM ethylene diamine tetra-acetic acid [EDTA], heparin 0.1 mg/l, pH 7.4). Following centrifugation, cells were resuspended in 80 µl of labeling buffer (PBS supplemented with 0.5% BSA, 2mM EDTA, pH 7.2) along with 20 µl of CD144 microbeads (Cat# 130-097-857 Miltenyi Biotec, Birgisch Gladbach, Germany). Cells were incubated for 15 min at 4 deg C. After incubation, cells were sorted through a column in a magnetic field (QuadraMACS, Miltenyi). Cells were washed 3 times with labeling buffer. Labeled ECs were recovered by gravity flow with 4 ml of cold labeling buffer after removing the column from the magnet. Isolated cells were quickly pelleted by centrifugation (400 x g for 5 min at 4 deg C with no brake) and immediately placed in RNA lysis buffer per protocol (Qiagen, RNeasy Plus Micro, Venlo, the Netherlands).RNA sequencing was performed on Illumina HiSeq 2500 instrument. Sequencing reads were mapped in a splice-aware fashion to the human reference transcriptome (hg19/GRCh37.75 assembly) using STAR aligner. Read counts over transcripts were calculated using HTSeq based on the Ensembl annotation for hg19/GRCh37.75 assembly. For differential expression analysis, we examined genes where at least 9 samples had >1 counts/million reads mapped and used the EdgeR method to generate log₂-fold change values.