characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-06-20T00:20:01Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-06-20T00:20:01Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112138262
characteristics.characteristic: External Id
characteristics.text: ArrayExpress
characteristics.characteristic: INSDC center name
characteristics.text: 2023-06-20T00:20:01Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-06-20T00:20:01Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14247571
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12436:HL60_Ewa_B3D5_WT
characteristics.characteristic: Submitter Id
characteristics.text: 36
characteristics.unit: year
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: HL-60
characteristics.characteristic: cell_line
characteristics.text: promyeloblast
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: HL-60 cells were grown in RPMI 1640 media with 10% heat-inactivated FBS (#10500-064, Gibco) and 1% Penicillin-Streptomycin (#15140-122, Gibco) in a humidified incubator at 37°C with 5% CO2. Cells were differentiated by adding 1.5% DMSO (#D2438, Sigma Aldrich). Each independently-differentiated batch was treated as a biological replicate. Cells were harvested before and at 24, 48, 72, 96 and 120 hours after differentiation with medium supplementation of 1.5% DMSO (#D2438, Sigma Aldrich). CRISPR/Cas9 generation in HL-60 cells was performed as described previously (Graziano et al. PLoS Biol 2019). Cloning of the target guide sequence to target Snx33 was performed using CRISPR/Cas9 technology. (Forward: CACCGctgggacgacGGATGCACAG; Reverse: aaacCTGTGCATCCgtcgtcccagC). Cells expressing BFP-tagged Cas9 were single-cell sorted in 96-well plate on BD FACSAria™ Fusion at EMBL Flow Cytometry Core Facility. Single-cell clones were verified by genomic DNA amplification by Touchdown PCR and sequencing, followed by Western blot of selected clonal lines. Total RNA samples obtained from 3 biological replicates were purified using RNeasy Mini Kit (#74104, Qiagen) according the manufacturer instructions with a DNase digestion step (#79254, Qiagen) using 10 mln of cells for each sample preparation Total RNA samples obtained from 3 biological replicates were purified using RNeasy Mini Kit (#74104, Qiagen) according the manufacturer instructions with a DNase digestion step (#79254, Qiagen) using 10 mln of cells for each sample preparation. To ensure high quality, samples were analyzed on an Agilent 2100 Bioanalyzer (Agilent Technologies).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: acute promyelocytic leukemia
characteristics.characteristic: disease
characteristics.text: none
characteristics.characteristic: genetic modification
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: blood
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: HL60_Ewa_B3D5_WT
characteristics.characteristic: title