Supplementary Materials from <i>apterous A</i> specifies dorsal wing patterns and sexual traits in butterflies

Butterflies have evolved different colour patterns on their dorsal and ventral wing surfaces to serve different signalling functions, yet the developmental mechanisms controlling surface-specific patterning are still unknown. Here, we mutate both copies of the transcription factor <i>apterous</i> in <i>Bicyclus anynana</i> butterflies using CRISPR/Cas9 and show that <i>apterous A,</i> expressed dorsally, functions both as a repressor and modifier of ventral wing colour patterns, as well as a promoter of dorsal sexual ornaments in males. We propose that the surface-specific diversification of wing patterns in butterflies proceeded via the co-option of <i>apterous A</i> or its downstream effectors into various gene regulatory networks involved in the differentiation of discrete wing traits. Further, interactions between <i>apterous</i> and sex-specific factors such as <i>doublesex</i> may have contributed to the origin of sexually dimorphic surface-specific patterns. Finally, we discuss the evolution of eyespot number diversity in the family Nymphalidae within the context of developmental constraints due to <i>apterous</i> regulation