Es cell qualified fbs

Cardiomyocytes from newborn rats was prepared by collagenase II digestion and cultured in DMEM/M199 medium with 10% FBS, 100 U/mL of penicillin, 100 μg/mL of streptomycin and 0.1 mM of Brdu (Sigma, St. Louis, MO, USA). C-kit(+) CSCs were purified by staining with PE-conjugated anti-c-kit monoclonal (#553355, BD) and sorting with a FACS Aria high-speed cell sorter (Becton Dickinson, San Jose, CA, USA), cultured in DMEM-Ham’s F-12 containing 10 ng/mL of EGF (PeproTech, Princeton, NJ, USA), 20 ng/mL of bFGF (PeproTech), 10 ng/mL of LIF (PeproTech), 10% of ES-cell qualified FBS (Gibco, Carlsbad, CA, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL). The cardio fibroblasts were removed from cardiomyocytes through differential adhesion.

Mouse embryonic stem (mES) cells, wildtype (wt) E14 (provided by Dr. Zhou-Feng Chen and Dr. Helle Færk Jørgensen) and Eed^−/− (provided by Dr. Anton Wutz) were cultured on 0.1% (w/v) gelatin-coated plates in ES medium (Glasgow Minimum Essential Medium (Sigma) supplemented with Glutamax-1 (Gibco), non-essential amino acids (Gibco), 50 mM 2-mercaptoethanol, 15% (v/v) ES-cell-qualified FBS (Gibco), and 1% (v/v) penicillin/streptomycin) in the presence of 1,000 U/ml of LIF (Millipore). To induce histone phosphorylation, the mES cells were stimulated with 1 µg/mL anisomycin in DMSO or DMSO only as control.
For ChIP, cells were cross-linked for 10 min at room temperature in culture media containing 1% formaldehyde, 10 mM Hepes (pH 8.0), 0.1 mM EGTA, and 20 mM NaCl. Cross-linking was stopped by addition of glycine to a final concentration of 0.125 M, followed by an additional incubation for 5 min. Fixed cells were washed 3 times with PBS and harvested in SDS lysis buffer (50 mM Tris at pH 8.1, 0.5% SDS, 100 mM NaCl, 5 mM EDTA, 1 mM PMSF, 10 µg/ml leupeptin and 10 µg/ml aprotinin). The cells were then pelleted for 10 min at 2,400 g followed by the same ChIP protocol as for striatal tissue. The included primer sequences are listed in Table S3.

For molecular cloning, all restriction enzymes, T4 DNA polymerase and T4 DNA ligase are from NEB (Ipswich, MA, USA). For mammalian cell culture, DMEM, common FBS, ES cell qualified FBS are from Invitrogen (Carlsbad, CA, USA). Antibiotics used for stable cell selection are from Invitrogen and Sigma (St Louis, MO, USA). CCE cells [18] (link), [19] (link), a mouse ES cell line, were a gift from Stem Cell Technologies (Vancouver, BC, Canada). The mAmetrine and tdTomato FPs are subcloned from Addgene plasmid 18879 [20] (link). All other FPs are from Clontech (Mountain View, CA, USA).