Quantitative Analysis of Protein Interactions

All data obtained from BLI on Octet RED 96 system (ForteBio, Fremont, CA, USA) were processed in exact accordance to the validation of this assay [32 (link)]. The association step data (in triplicate) were obtained in an excel.csv file extracted from raw outputs of the Octet RED 96 system and then imported into Prism 7.0 software (GraphPad Software Inc., La Jolla, CA, USA) where area under the curve (AUC) calculations were made and graphs produced.
Phylogenetic trees were obtained from timetree.org and then further manually recreated using Mesquite software version 3.2 (http://mesquiteproject.org/). The obtained phylogenetic trees were then further analysed in RStudio (R Core Team, 2015) for all comparative analysis using the Ape package [61 (link)]. Heat-mapping of AUC values over the phylogenetic trees was achieved using the contMap function of the R package phytools [62 (link)].