The mRNA-Seq data of human-Zic3 overexpression in chicken limbs

We observed Zic3 highly expressed in the anterior regions of limb buds in all investigated tetrapods. To investigate the function of the Zic3 in limbs, we overexpressed the human-Zic3 gene in the lateral plate mesoderm (LPM) of developing chicken hindlimbs at HH10 using the RCAS avian retrovirus. Subsequently, the embryos were allowed to incubate until HH30. Autopod and zeugopod regions were dissected and subjected to transcriptome sequencing (mRNA-Seq), with the corresponding positions from individuals injected with the empty RCAS vector serving as controls. The results of the differential analysis and enrichment analysis revealed the regulatory network of Zic3 and various regulatory pathways that may be affected. After 5 days of infection (HH30), the entire autopod and zeugopod regions of individuals injected with RCAS-Zic3 were collected under a microscope. The same procedure was applied to the control group, where the corresponding areas of individuals injected with RCAS were collected. Each group included five biological replicates.