Chromatin immunoprecipitation analysis (ChIP), gene expression analysis by RT-PCR, protein co-immunoprecipitation assays and western blot analysis were performed as previously described (Gomar-Alba et al., 2017 (link)). In ChIP assays, Dynabeads Protein G magnetic beads were incubated with HA-probe (F-7) antibody (Santa Cruz Biotechnology; SC-7392) or monoclonal anti-GFP (Roche Diagnostics; 11814460001). The primary antibodies used in the western blot analysis were monoclonal anti-HA peroxidase 3F10 antibody (Roche Diagnostics; 12013819001) diluted 1:5000, monoclonal anti-GFP (Roche Diagnostics; 11814460001) diluted 1:5000 and monoclonal anti Cdc2 p34 (PSTAIRE; Santa Cruz Biotechnology; SC-53) diluted 1:2000. Blots were developed using anti-mouse IgG and anti-rabbit IgG horseradish peroxidase-conjugated secondary antibodies (1:20,000; Thermo Fisher Scientific; 170-6516 and 31460, respectively) and Supersignal West Femto Maximum Sensitivity substrate (Thermo Fisher Scientific). Bands were quantified using an ImageQuant LAS 4000mini Biomolecular Imager (GE Healthcare).
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