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.