organism: Homo sapiens
description: Protocols: Normal human epidermal keratinocytes (NHEKs) were isolated from foreskin sample of young donors (less than 7 years) as described (Gilchrest, 1983, doi: 10.1111/1523-1747.ep12541084). Samples were obtained after circumcision following approval by the ethic committee of the Clinique St Luc (Bouge, Belgium). NHEKs were grown in specific keratinocyte serum-free medium (K-SFM) (Invitrogen, 15005042), supplemented with bovine pituitary extract (50 µg/ml) and human recombinant epidermal growth factor (5 ng/ml). For UVB-SIPS model, early passage NHEKs were seeded at low confluence in K-SFM. At 72 h after plating, cells were exposed to UVB radiation through a thin layer of PBS. UVB (NB-UVB) used are generated by a Narrow-Band UVB lamp (Philips, TL20W/01 lamps) with a peak at 311 nm. The emitted radiation was checked under the flask lid using a UVB radiometer with a UVB sensor (Bioblock Scientific). After irradiation, PBS was replaced by K-SFM medium. UVB exposures are repeated three times on a single day with a three-hours interval. Unexposed cells (controls) were kept in the same culture conditions without UVB exposure. Total RNA was extracted using RNeasy mini kit (Qiagen) according to manufacturer's instructions. RNA integrity was checked with Agilent Bioanalyzer. Sequencing mRNA libraries were prepared using TruSeq® Stranded mRNA library preparation kit (Illumina) with 1 µg of total RNA by condition.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-12-31
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-12-31
attributes.tag: External Id
attributes.value: SAMEA10160857
attributes.tag: INSDC center alias
attributes.value: University of Namur
attributes.tag: INSDC center name
attributes.value: University of Namur
attributes.tag: INSDC first public
attributes.value: 2022-12-31T00:57:08Z
attributes.tag: INSDC last update
attributes.value: 2022-12-31T00:57:08Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10969:KC20_CTL3J
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: keratinocyte
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: juvenile stage
attributes.tag: disease
attributes.value: normal
attributes.tag: environmental history
attributes.value: replicating cells
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: 1
attributes.tag: irradiate
attributes.value: none
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: skin of prepuce of penis
attributes.tag: passage
attributes.value: 4 to 5
attributes.tag: sample name
attributes.value: E-MTAB-10969:KC20_CTL3J
attributes.tag: sampling time point
attributes.value: 3
attributes.tag: sex
attributes.value: male