title: Illumina HiSeq 1500 sequencing; Ablation of casein kinase 1α in keratinocytes induces p53-dependent hyperpigmentation of the skin
design: Ablation of casein kinase 1α in keratinocytes induces p53-dependent hyperpigmentation of the skin
library_construction_protocol: Mouse ear dorsal-epidermis and tail skin were homogenized in QIAzol Lysis Reagent (Qiagen, Hilden, Germany) with a TissueLyser (Qiagen) according to the manufacturer’s protocol. Tail and ear cells were pooled. Csnk1a1 floxed and Csnk1a1/p53 double-floxed mice were reported before.14 K14–Cre–ERT2 mice, purchased from the Jackson Lab (Bar Harbor, ME, USA) were used as a skin-specific deleter strain. Csnk1a1△skin and Csnk1a1/p53△skin mice were developed by crossing Csnk1a1 floxed or Csnk1a1/p53 floxed mice with K14–Cre–ERT2 mice. Mice were bred at the National Laboratory Animal Center, Tainan, Taiwan, and were maintained at an SPF animal facility at China Medical University. CK1a knock-out and CK1a/p53 double knock-out mice where treated topically with 4-hydroxytamoxifen (4HT; H6278, Sigma) dissolved in 99.9% alcohol (32205, Sigma) everyday for 2 weeks (1.5mg/0.5ml). To compare the effect of CK1a ablation in the epidermis with UV exposure, UVB mice where exposed to UV irradiation (150 mJ/cm^2) every other day for 2 weeks. Epidermis was taken at different time points for transcriptome analysis (7 and 14 days). Following chloroform extraction, ethanol precipitation and DNase digestion, RNA was purified using an RNeasy Mini Kit (Qiagen). RNASeq libraries were prepared using a TruSeq Stranded mRNA Sample Prep kit (Illumina, Tokyo, Japan).
attributes.tag: Experimental Factor: K14-Cre-ERT2 / CK1 fl/fl / p53 fl/fl
attributes.value: genotype
attributes.tag: Experimental Factor: 4-hydroxytamoxifen
attributes.value: compound
attributes.tag: Experimental Factor: none
attributes.value: irradiate
attributes.tag: Experimental Factor: 14
attributes.value: time