biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsTomoko Hashimoto-Tamaokitranscription profiling by arrayHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-GEOD-14838In the epidermis, keratinocytes are involved in physical and first-line immune protection of the host. In this work, we analyzed molecular responses after the addition of contact sensitizers, 2,4-dinitrochlorobenzene (DNCB) using cultured human keratinocytes from the bulge region of a plucked hair follicle (bulge-derived keratinocytes; BDKs) in comparison with neonatal human epidermal keratinocytes NHEK and human monocytic leukemia THP-1. The changes in gene expression by the treatment of DNCB in BDKs were different from those in THP-1. Many genes orchestrating keratinocyte differentiation, which interact TGF-β and BMP signaling pathway, were significantly up-regulated in response to DNCB. Keywords: Comparison analysis of gene expression changes by the treatment of DNCB three cell strains composed of four samples; two BDKs established from different donors, NHEK, and THP-1. One replicate per array.biostudies-arrayexpressGrowth Protocol - BDKs and NHEK ; cultured in Defined Keratinocyte-SFM medium （DK-SFM, GIBCO/Invitrogen Co., CA） at 37°C in a humidified atmosphere containing 5% CO2. THP-1; cultured in RPMI1640 medium (GIBCO/Invitrogen Co.) containing 10% fetal bovine serum (Equitech-bio Inc., TX, USA) at 37°C in a humidified atmosphere containing 5% CO2.Sample Processing - BDKs or NHEK ; seeded at 5 x 105 cells/60 mm φ dish and incubated in medium at 37 °C for 24 hr, and sequentially in MCDB153 medium (Research Institute for the Functional Peptides Co., Ltd, Yamagata, Japan) at 37 °C for 24 hr. 24 hr later, cells were treated with 10 μM DNCB or 0.1% DMSO for 4.5 hr. THP-1 ; seeded at the density of 7 x 105 cells/10 mm φ dish. 24 hr later, cells were treated with 10 μM DNCB or 0.1% DMSO for 4.5 hr.Labeling - Total RNAs (each 200ng) were labeled with Cy3- or Cy5- CTP using Agilent Quick Amp Labeling Kit (Agilent Technologies Inc.).Nucleic Acid Extraction - Isolated using TRIzol reagent (Invitrogen, CA, USA) and purified using an RNeasy kit (QIAGEN, Hilden, Germany).Hybridization - Cy3- or Cy5-labeled probes were mixed, fragmented, and then dissolved in GE hybridization buffer in the Gene Expression Hybridization Kit (Agilent). After hybridization, slides were washed sequential.MIAME ScoreRaw DataOrganizationAssays and DataMAGE-TAB FilesArray DesignsYoshie YoshikawaTomoko Hashimoto-TamaokiYukio KitanoHiroki ShimaNozomi KanazawaYusuke SasaharaAssay Data Transformation - ID_REF = <br>VALUE = normalized log2 ratio (Cy5/Cy3) representing test/referenceImage Adquisition - Scanned on an Agilent microarray scanner.Image Adquisition - Images were quantified using Agilent Feature Extraction Software (ver9.5.3.1).Feature Extraction - LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Expressionist (GeneData AG, Basel, Switzerland) software was used.falseGene expression changes by 2,4-dinitrochlorobenzene in human bulge-derived keratinocytesIn the epidermis, keratinocytes are involved in physical and first-line immune protection of the host. In this work, we analyzed molecular responses after the addition of contact sensitizers, 2,4-dinitrochlorobenzene (DNCB) using cultured human keratinocytes from the bulge region of a plucked hair follicle (bulge-derived keratinocytes; BDKs) in comparison with neonatal human epidermal keratinocytes NHEK and human monocytic leukemia THP-1. The changes in gene expression by the treatment of DNCB in BDKs were different from those in THP-1. Many genes orchestrating keratinocyte differentiation, which interact TGF-β and BMP signaling pathway, were significantly up-regulated in response to DNCB. Keywords: Comparison analysis of gene expression changes by the treatment of DNCB three cell strains composed of four samples; two BDKs established from different donors, NHEK, and THP-1. One replicate per array.2010-04-09T00:00:00Z2023-08-05T08:40:54.295Z2022-03-03T15:29:05.362ZE-GEOD-14838GSE14838EFO_0002768