Bioruptor ucd 200 ultrasound sonicator

ChIP was performed using the ChIP-IT express kit (Active Motif). Briefly, HUVEC transfected with ERG or control siRNA, and/or treated with Ang1, were crosslinked for 10 min with formaldehyde (to a final concentration of 1%). Chromatin was sheared for five cycles (30 s on, 30 s off) using a Bioruptor UCD-200 ultrasound sonicator (Diagenode), resulting in DNA fragments of 200–1,000 bp in size. Chromatin was immunoprecipitated with 2 μg antibody to ERG (sc-354X, Santa Cruz Biotechnology), or negative control rabbit IgG (PP64, Chemicon, Millipore). Immunoprecipitated DNA was then used as template for quantitative PCR using primers specific for genomic loci. Oligonucleotide sequences are listed in Supplementary Table 1.

Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT Express kit (Active Motif)^{33 (link)}, according to the manufacturer’s instructions. In brief, HUVEC were cross-linked for 10 min with 1% formaldehyde. Chromatin was sheared using a Bioruptor UCD-200 ultrasound sonicator (Diagenode) and immunoprecipitated with 3 μg antibody to ERG (sc-354X, Santa Cruz Biotechnology), KLF2 (rabbit, ab203591, Abcam), H3K27Ac (rabbit, 39133, Active Motif) or p300 (mouse, ab14984, Abcam). The respective negative controls were rabbit IgG (PP64, Chemicon, Millipore) and mouse IgG (12- 371, Millipore). Immunoprecipitated DNA was then used as template for quantitative PCR using primers specific for genomic loci and listed in Supplementary Table 5.

Chromatin immunoprecipitation (ChIP) was carried out using ChIP-IT express kit (Active Motif). HUVEC were transfected with ERG or control siRNA (20 nM, for 48 h) or treated with PBS or TGFβ2 (10 ng ml⁻¹, for 30 min), and cross-linked for 10 min with formaldehyde (final concentration of 1%). Chromatin was sheared using a Bioruptor UCD-200 ultrasound sonicator (Diagenode), resulting in DNA fragments of 500–1000 bp in size. Chromatin was immunoprecipitated with 2 μg antibody to ERG (sc-354X, Santa Cruz Biotechnology), SMAD3 (#9523, Cell Signalling) or negative control rabbit IgG (PP64, Chemicon, Millipore) using protein G magnetic beads (Active Motif). Immunoprecipitated DNA was then used as template for qPCR using primers specific for genomic loci. Oligonucleotide sequences are listed in Supplementary Table 5.