ChIP–western blot

For western blot analysis of chromatin immunoprecipitation, after washing the immunoprecipitates, 4 × reducing SDS Loading Dye (500 mM Tris HCl, pH 6.8, 8% SDS, 40% glycerine, 20% β-mercaptoethanol, 5 mg ml⁻¹ bromophenol blue) was added to the beads as well as to the lysates and the supernatants and samples were incubated at 99 °C for 20 min and loaded onto SDS–PAGE, proteins were then transferred into PVDF membrane for western blotting analyses with anti Tbx1 antibody (Abcam, #ab18530, 1:1,000 or LifeSpan Biosciences LS-{"type":"entrez-nucleotide","attrs":{"text":"C31179","term_id":"2362975","term_text":"C31179"}}C31179/7296, 1:500), anti-SETD7 antibody (ab124708, 1:500), anti MLL1 Antibody (Bethyl Laboratories, A300-074A, 1:1,000), anti-KMT2C/MLL3 antibody (ab71200, 1:500), anti-KMT2B/MLL4 antibody (ab104444; 1:500) and Amersham ECL Rabbit IgG, HRP-linked whole Ab (GE Heathcare, #NA934V; 1:10,000) as secondary antibody.