Fig. 5

Injury-induced emergence of RPCs in the mouse retina following Prox1 deletion in BCs.

a Single-cell RNA sequencing (scRNA-seq) analysis was performed to examine mRNA expression in cells from Prox1^fg/fg and Prox1^fg/fg;Chx10-CreERT2 mouse retinas before and after MNU injury (see “Methods” for details). Uniform Manifold Approximation and Projection (UMAP) plots show retinal cells clustered by cell type. Red arrowheads indicate cluster #20. b The identity of each cell cluster is summarized in the accompanying table. c RNA velocity profiles derived from scRNA-seq data for MNU-injured Prox1^fg/fg and Prox1^fg/fg;Chx10-CreERT2 mouse retinas. Magnified views of boxed areas in the top panels are displayed in the bottom. d The frequency of cluster #20 in the retinas of the indicated mouse genotypes is shown in the graph. e Violin plots depicting the expression levels of Gadd45a, Hbegf, Notch1, and Hes1 mRNA in the indicated cell clusters identified through scRNA-seq analyses of MNU-injured Prox1^fg/fg and Prox1^fg/fg;Chx10-CreERT2 mice. Each dot represents gene expression level in individual cells, with the number of dots (n) indicated for each group. Blue horizontal bars indicate mean expression values. f, g The distribution of Hes1 in MNU-injured Prox1^fg/fg and Prox1^fg/fg;Chx10-CreERT2 mice mouse retinas was analyzed by immunostaining (images provided in Supplementary Fig. 14a). Graphs show the number of Hes1-expressing MG (f) and the intensity of Hes1 in MG (g). The number of retinas analyzed is indicated in the graphs. h Violin plots illustrate the expression levels of E2f5, Pcna, Cdk4, and Ccnd1 mRNA in specific cell clusters. i, j The distribution of Ccnd1 in retinas was analyzed by immunostaining (images shown in Supplementary Fig. 14b). Graphs display the number of Ccnd1-expressing MG (i) and the intensity of Ccnd1 in MG (j). Error bars denote SEM. P-values were calculated using one-sided Student’s t-test (*, p < 0.05; ****, p < 0.001).