2.3 Secondary structure and conservation prediction
Secondary structure prediction of full-length IRX4 protein was performed using the PSIPRED server (http://bioinf.cs.ucl.ac.uk/psipred/)41. ConSurf server was used to estimate and visualize the evolutionary conservation of residues in the homeobox region of IRX4 (https://consurfdb.tau.ac.il/)42. The maximum number of homologs for conservation analysis was kept at 150, and the E-value cut-off of 0.0001. The phylogenetic tree was constructed using neighbour-joining algorithm.