characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-09-30T17:01:58Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-12T11:50:01Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104679824
characteristics.characteristic: External Id
characteristics.text: Section of Bioinformatics and Systems Cardiology, Department of Internal Medicine III and Klaus Tschira Institute for Integrative Computational Cardiology, University of Heidelberg, 69120 Heidelberg, Germany DZHK (German Centre for Cardiovascular Research), Partner site Heidelberg/Mannheim, 69120 Heidelberg, Germany
characteristics.characteristic: INSDC center name
characteristics.text: 2018-09-30T17:01:58Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-12T11:50:01Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS2281207
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-6564:49746
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: HeLa
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001185
characteristics.characteristic: cell line
characteristics.text: epithelial cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000066
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: HeLa Flp-In T-REx cells were cultured in DMEM supplemented with 9% fetal bovine serum and 1x Penicillin Streptomycin. The cells were cultivated at 37°C and 5% CO2 in a humidified incubator. For siRNA transfection, cells were reverse transfected with 60 pmol siRNA targeting RNPS1 (or Luc in the negative control) per 2 x 10^5 cells using 2.5 μl Lipofectamine RNAiMAX. For rescue, for generating stable RNPS1 rescue cell lines, the expression constructs were cloned into the pcDNA5/FRT/TO vector containing an N-terminal FLAG-tag. Standard protocols were used to generate stable rescue or reporter HeLa Flp-In T-REx cell lines and positive clones were selected with 100-150 μg/ml hygromycin B. Expression of stable cell lines was induced with 1 μg/ml doxycycline 24 h after siRNA transfection. Fresh medium with 1 μg/ml doxycycline was added after additional 48 h. The H176D-E179R mutation of a surface exposed patch of RNPS1 prevents ASAP complex formation. Total RNA was extracted with peqGOLD TriFast according to the manufacturer's instructions. Ribosomal depletion and strand specific library preparation was carried out with the TruSeq R Stranded Total RNA LT (with Ribo-ZeroTM Human/Mouse/Rat) according to the manufacturer's instructions.
characteristics.characteristic: description
characteristics.text: cervical adenocarcinoma
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001416
characteristics.characteristic: disease
characteristics.text: pcDNA5/FRT/TO-RNPS1
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: uterine cervix
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000002
characteristics.characteristic: organism part
characteristics.text: RNPS1 knockdown and rescue with wild type RNPS1
characteristics.characteristic: phenotype
characteristics.text: RNPS1 knockdown by siRNA
characteristics.characteristic: rna interference
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: 49746
characteristics.characteristic: title