NextSeq 500 sequencing; YAP-TEAD multiphasic activation regulates pluripotent stem cell mechanics to drive cardiac specification and contractility in pathological heart.
YAP-TEAD multiphasic activation regulates pluripotent stem cell mechanics to drive cardiac specification and contractility in pathological heart.
Undifferentiated iPSCs were seeded onto 100 mm dishes (four plates for each sample) in complete medium for 4 days. For cardiomyocytes, the cells were differentiated for 15 days and beating areas were enriched before being processed Chromatin was immunoprecipitated from three technical replicates using a ChIPgrade anti-YAP antibody (Cell Signaling Technologies, see Supplementary Table 8) and following the manufacturer's protocol (Pierce™ Agarose ChIP Kit, Thermo Fisher Scientific). A control ChIP was performed using rabbit immunoglobulins (IgG). The samples were eluted in 30 mL eluting buffer and stored at -80 °C before analysis. For library preparation, the size distribution of each ChIP DNA sample was assessed by running a 1 µL aliquot on Agilent High Sensitivity DNA chip using an Agilent Technologies 2100 Bioanalyzer (Agilent Technologies). The concentration of each DNA sample was determined using a high sensitivity Quant-iT™ dsDNA Assay Kit and a Qubit Fluorometer (Thermo Fisher Scientific). Purified ChIP DNA (10 µg) was used as the starting material for sequencing libraries preparation
Experimental Factor: immunoprecipitate
Anti YAP 15d