Setd1b

Co-Immunoprecipitation was performed according to the manufacturer’s protocol and the following kit was used: Dynabeads^® Protein G Immunoprecipitation Kit (Thermo Fischer, Waltham, USA; #10007D). Briefly, cell lysates (1000 µg per sample) were incubated with the primary antibodies over night at 4 °C with rotation. Following primary antibody was used: SETD1B (Abcam, Cambridge, UK; #ab113984; 10 µg per sample). The next day, the targets were eluted and stored for further use or directly analyzed by western blotting as recently described.

The PLA was performed according to the manufacturer’s manual and the following reagents were used: Duolink^® In Situ Detection Reagents Red (Sigma-Aldrich, Darmstadt, Germany; #DUO92008-100RXN), Duolink^® In Situ PLA^® Probe Anti-Mouse PLUS (Sigma-Aldrich, Darmstadt, Germany; #DUO92001-100RXN) and Duolink^® In Situ PLA^® Probe Anti-Rabbit MINUS (Sigma-Aldrich, Darmstadt, Germany; #DUO92005-100RXN). In short, the cells were co-stained over night at 4 °C using the following antibodies: SETD1B (Abcam, Cambridge, UK; #ab113984; 1:1000) and ZEB1 (R&D Systems, Wiesbaden, Germany; #639914; 10 µg/ml). The next day, the PLA probe solution was added to the cells as described in the protocol. After the ligation and amplification steps, the nuclei were stained using ProLong® Gold Antifade reagent (Life Technologies, Darmstadt, Germany). Detection was performed using a Nikon Eclipse Ti-S fluorescence microscope (Nikon, Tokyo, Japan).