Fig. 7: Hmx1 misexpression in mouse leads to widespread abnormal development in pinna structures including cartilage, fibroblasts, muscle, and epidermis.

a Diagram of micro-dissected pinna prominence at E14.5 for bulk and sc-RNA sequencing. b UMAP dimensional reduction visualizes seventeen cell clusters of pinna prominence at E14.5. c Hmx1 expression is mainly expressed in fibroblasts and expanded in these cells in mEC1^dup/dup compared with wild type. d RNA-seq analysis compares the whole pinna structure between wild-type and E14.5 mEC1^dup/dup mice. Differentially expressed genes (DEGs) are defined as FDR ≤ 0.05, log2 CPM ≥ 1, and |log2 FC | 0. Up- and down-regulated genes are shown as gold and blue dots, respectively. Important DEGs including two previously reported cartilage development-related genes including Comp⁸⁵ and Arhgap36⁸⁶ were labeled. e, f Single cell differentially gene analysis in chondrocytes (e) and epithelial cell (f) cluster. DEGs are defined as FDR value ≤ 0.05 and |log2 FC | 0. Up- and down-regulated genes are shown as red and blue dots, respectively. Some genes that are critical for cell development are labeled. Source data are provided as a Source Data file. g, Violin plots show gene expression comparison between wild type and mEC1^dup/dup in each cell cluster. Red rectangle represents genes that are significant changed in corresponding cell clusters (Hmx1 in fibroblast cluster, Msx1 in chondrocyte cluster, Krtdap and Dmkn in epithelial cell cluster). h, Masson’s trichrome staining at E14.5 (upper and lower pinna parts) is shown for WT and mEC1^dup/dup. Scale bars measure 100 µm. Experiments were performed with three independent embryos.