2024-03-13
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Cardiovascular Regeneration Centre of Molecular Medicine Goethe-University Frankfurt
INSDC center name
public
INSDC status
ERS18331590
SRA accession
E-MTAB-13833:Sample 1
Submitter Id
ArrayExpress
broker name
hPC-PL
cell_line
pericyte
cell_type
not collected
collection date
human
common name
Protocols: Human Placenta Pericytes (hPC-PL, C-12980, PromoCell) were seeded at 6000 cells/cm2 density and culture in the respective Pericyte Growth Medium 2 (C-28041, PromoCell). RGS5 silencing was performed using siRNA. RGS5 siRNA (ID: SASI_Hs01_00094781, Sigma-Aldrich) and control siRNA (sense: CGUACGCGGAAUACUUCGA; antisense: UCGAAGUAUUCCGCGUACG) were used at a final concentration of 50nM. We used hPC-PL from different donors (464Z005, 463Z019.1, 422Z033.3). Total RNA was extracted from human placenta hPC-PL using the RNeasy Plus Mini Kit (Qiagen) according to the manufacturer’s instructions. Total RNA was extracted from cells using the RNeasy Plus Mini Kit (Qiagen) according to the manufacturer’s instructions. The synthesis of complementary DNA (cDNA) was performed by the reverse transcription (RT) of mRNA using M-MLV reverse transcriptase (Thermo-Fisher Scientific). Quantitative PCR reactions were performed using the StepOnePlus real-time PCR cycler (Thermo-Fisher Scientific) and relative gene expression was calculated by normalisation to ACTB gene expression (2-Ct). Human primers used: RGS5 (f: AGAAACCAGCCAAGACCCA, r: GGAGTTTGTCCAGGGAATCAC), ACTB (f: CATGTACGTTGCTATCCAGGC, r: CTCCTTAATGTCACGCACGAT), ACTA2 (f: CATCATGCGTCTGGATCTGG, r: GACAATCTCACGCTCAGCAG), TGFB2 (f: AAGAAGCGTGCTTTGGATGCGG, r: ATGCTCCAGCACAGAAGTTGGC). Total RNA was used as input for whole RNA-Seq library preparation, for which integrity was verified with LabChip Gx Touch 24 (CLS138162, Perkin Elmer). 1 µg of total RNA was used as input for Truseq Stranded total RNA library preparation following the low sample protocol (Illumina).
description
neonate
developmental stage
none
disease
siControl
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
placenta
organism part
Homo sapiens
scientific_name
Sample 1
title