Figure 2. Ventral ependymal progenitors undergo EMT after SCI.

(A) Expression of EMT-related genes by FACS-seq. Heatmap representation of epithelial and mesenchymal gene expression in ctgfa⁺gfap⁺ cells. Color scale in the dendrogram represents log₂(fold enrichment) in ctgfa⁺gfap⁺ cells relative to control SCs.

(B) Enrichment of EMT markers by FACS-seq. Log₂(fold enrichment) was averaged for epithelial and mesenchymal genes.

(C) Distribution of EMT markers by FACS-seq. Upregulated genes were subdivided into log₂(FE) >1 (Green) and log₂(FE) = 0 to 1 (Light green). Downregulated genes were subdivided into log₂(FE) = −1 to 0 (light red) and log₂(FE) < −1 (red). Percent genes within each category are shown.

(D) RNAscope for twist1a in wild-type SCs at 1 and 2 wpi, and in uninjured controls. Hematoxylin staining (blue) was used as a counterstain. At 1 wpi, SC cross sections at the lesion site and distal to the lesion from the rostral side are shown. Arrowheads point to twist1a expression at the lesion core at 1 wpi. Arrows point to twist1a expression in ventral ependymal progenitors at 1 and 2 wpi.

(E–H) Cdh2 (E) and Cdh1 (G) immunostaining in ctgfa:EGFP zebrafish at 1 and 2 wpi, and in uninjured controls. SC cross sections are shown, and dotted lines delineate central canal edges. Arrowheads point to domains of co-expression of Cdh2 and EGFP (E), and domains of diminished Cdh1 expression and increased EGFP expression (G). Quantification of Cdh2 (F) and Cdh1 (H) expression in ctgfa- and ctgfa⁺ cells at 2 wpi are shown. ****p < 0.0001. Scale bars, 50 μm.