mESC_FKBP12(F36V)-Zscan10_cloneN2_DMSO
Protocols: 1 million mESCs were detached with Accutase (Innovative Cell Technologies). Mouse embryonic stem cells (mESCs) were cultured in 2i+LIF medium composed of DMEM (Thermo Fisher, 10566-024), 10% ES Cell FBS (Thermo Fisher, 16141079), 1X MEM Non-Essential Amino Acids Solution (Thermo Fisher, 11140050), 1 mM sodium pyruvate (Thermo Fisher, 11360-039), 100 µM 2-Mercaptoethanol (Thermo Fisher, 31350-010), 100 U/ml Penicillin-Streptomycin (Thermo Fisher, 15140-122), 100 ng/ml of mouse LIF (Protein Production and Structure Core Facility, EPFL), 3 μM of GSK3 inhibitor (CHIR99021; Tocris Biosciences, 4423), and 2 μM of MEK1/2 inhibitor (PD0325901; Tocris Biosciences, 4192). To induce the degradation of Zscan10 endogenously tagged with an FKBP12(F36V), the cells were cultured in medium containing either 500 nM dTAGv-1 (MedChemExpress, HY-145514) or the same volume of DMSO for 72 h prior to harvest. RNA was extracted with High Pure RNA Isolation Kit (Roche, 11828665001). 500 ng of the isolated RNA was used to prepare RNA-seq libraries using QuantSeq 3′ mRNA-Seq V2 Library Prep Kit (Lexogen, 191.24) following the manufacturer's instruction.
isolate
not applicable
organism
Mus musculus
organism
Mus musculus
cell_line
ES cell line
collection date
not collected
scientific_name
Mus musculus
common name
house mouse
geographic location (country and/or sea)
not collected