In this study we described the protein-protein interaction network of the Drosophila Speciation Core Complex by analysing the interactome of its subunit: HMR, LHR, NLP, BOH1 (CG33213), BOH2 (CG4788) and HP1a. For this purpose we performed Affinity Purification coupled with Mass Spectrometry (AP-MS) in D. melanogaster SL2 cells using as bait the two hybrid incompatibility proteins HMR (n = 8) and LHR (n = 4), as well as NLP (n = 3), BOH1(CG33213, n = 4), BOH2 (CG4788, n = 5) and HP1a (n = 4). Each bait was targeted with at least one antibody (rat anti-LHR 12F4, mouse anti-HP1a 2C09, rabbit anti-Nlp, anti-FLAG-M2 for FLAG-CG33213 and FLAG-CG4788), while HMR was targeted with three different antibodies (rat anti-HMR 2C10 and 12F1, anti-FLAG-M2 for FLAG-HMR). Individual replicates and antibodies used are listed in samples_table.