Sort3_IFEpos
Mus musculus
Protocols: Keratinocytes were isolated from three-week-old Lgr6-EGFP-CreETR2 mice as described previously (Jaks et al., 2008). Briefly, dorsal skin was harvested, cut in strips and digested in trypsin. Epidermal cells were peeled off the dermis and singularized by magnetic stirring. Cells from 2 - 3 mice were pooled per experiment. The collected cells were stained with AlexaFluor647-conjugated anti-CD49f antibody and PE-Cy7-conjugated anti-Sca-1 antibody (both from BD Biosciences), and filtered before sorting. Keratinocytes stained for CD49f, Sca-1 and 7-AAD were live sorted using a FACSAria (BD). Lgr6+ IFE cells were defined as the CD49fhigh, Sca-1high and EGFPhigh fraction. Lgr6+ HF cells were defined as the CD49fhigh+dim, Sca-1neg and EGFPhigh fraction. One hundred thousand cells were collected for each population. RNA was isolated from sorted keratinocytes by combining the RNA-containing aqueous phase extract using RNA-Bee (Amsbio) with the RNAaqueous-Micro Kit (Ambion). RNA integrity was measured using an Agilent 2100 Bioanalyzer Pico Chip or a 2200 TapeStation High Sensitivity R6K Screen Tape. The RNA integrity number of the used samples was higher than 8.5. The sequencing library was generated according to the STRT protocol, described in (Islam et al., 2012), using 1 ng total RNA as input for each sample. For each sample two technical replicates were sequenced (suffix _1 and _2). Islam, S., Kjällquist, U., Moliner, A., Zajac, P., Fan, J.B., Lönnerberg, P. and Linnarsson, S. (2012). Highly multiplexed and strand-specific single-cell RNA 5' end sequencing. Nat Protoc 7, 813-828.
ENA-FIRST-PUBLIC
2015-10-22T17:01:25Z
ENA-LAST-UPDATE
2018-03-09T00:03:23Z
External Id
SAMEA3367440
INSDC center name
Karolinska Institutet Department of Biosciences and Nutrition
INSDC first public
2015-10-22T17:01:25Z
INSDC last update
2018-03-09T00:03:23Z
INSDC status
public
Submitter Id
E-MTAB-3533:Sort3_IFEpos
age
21 day
broker name
ArrayExpress
cell type
Lgr6-positive basal interfollicular epidermis cell
common name
house mouse
genotype
Lgr6cre-EGFP-IRES-CreERT2 heterozygous
organism part
dorsal skin
phenotype
CD49f hi/Sca1 hi/EGFP hi
sample name
E-MTAB-3533:Sort3_IFEpos
scientific_name
Mus musculus
sex
male
strain
C57BL/6