ChIP-Atlas: SRX4495313

Antigen

Cell type Class: Epidermis
Cell type: Hair Follicle
MeSH Description: A tube-like invagination of the EPIDERMIS from which the hair shaft develops and into which SEBACEOUS GLANDS open. The hair follicle is lined by a cellular inner and outer root sheath of epidermal origin and is invested with a fibrous sheath derived from the dermis. (Stedman, 26th ed) Follicles of very long hairs extend into the subcutaneous layer of tissue under the SKIN.

source_name: Hair Follicle Stem Cells
strain: C57BL/6
tisse: hair follicle
chip antibody: home-made lab (Chiacchiera, Rossi, Sriganesh Jammula, et al. 2016)
genotype: WT

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: nagen synchronized hair follicles from LGR5-Ring1a/b ++ mice were extracted following the protocol previously described. Single cells were resuspended in Sorting medium (as previously described) and LGR5-GFP positive cells were FACS-sorted with either FACS-Melody (‎BD FACSMelody Cell Sorter) or FACS-Jazz (‎BD FACSJAZZ Cell Sorter), discriminating only singlets (plotting Area vs Height on both SSC and FSC) and only alive cells with Propidium Iodide. An amount of 2.5 Milion cells were resuspended and sonicated with Branson Digital Sonifier (Branson) in IP buffer (SDS Buffer: Triton Buffer 2:1 – SDS Buffer (NaCl 100mM, Tris-HCl pH 8.1 50mM, NaN3 0.2%, SDS 0.5%); Triton Buffer ( NaCl 100mM, Tris-HCl pH 8.6 100mM, EDTA 5mM, NaN3 0.2%, Triton X-100 5%). Sonicated chromatin were incubated with 10ug of Ring1b antibody generated in the lab (Chiacchiera, Rossi, Sriganesh Jammula, et al. 2016) over night and the immunocomplexes were recovered by magnetic beads (Dynabeds, Life-technologies) and decrosslinked over night with NaHCO3 0.1M and 1% SDS. Decrosslinked DNA were purified.