We used RELAX (28), a hypothesis-testing framework, to assess whether the dsx sequences in certain monophyletic lineages of biological interest had experienced relaxed or intensified selection. RELAX is useful for identifying trends and/or shifts in the stringency of natural selection on a given gene. RELAX estimates a relaxation/intensification parameter (K). K > 1 in test lineages indicates intensified positive selection whereas K < 1 indicates relaxed selection. We used HYPHY 2.3.14 (37) for this estimation.

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