Fig. 5

Cervical epithelium differentiation reconstruction by single-cell clone culture. a Representative phase-contrast microscopic image of GFP+ mouse CVSC single-cell colonies. Scale bar, 100 μm. Mouse CVSC single-cell colonies stained with proliferative marker KI67, and adult epithelial stem cell markers KRT5, KRT17, and P63. Data representative of n = 3 biological replicates. Scale bar, 100 μm. b 3D differentiated single-cell colonies A and B at 2, 4, 6 and 8 days since 3D culture obtained for bulk RNA sequencing. Scale bars, 100 mm. c The heatmap showed the Pearson correlation coefficient between single-cell colony A and bulk RNA sequence data. Each row represents one cluster from single-cell RNA sequence data and each column represents one single cell clone-specific bulk RNA-seq sample (left). Schematic shows alignment of single-cell colony A bulk RNA-seq data from 5 in vitro time points projected onto the pseudo-time differentiation map. The alignment of bulk RNA-seq data is shown in the red line (right). d The heatmap showed the Pearson correlation coefficient between single-cell colony B and bulk RNA sequence data. Each row represents one cluster from single-cell RNA sequence data and each column represents one single cell clone-specific bulk RNA-seq sample (left). Schematic shows alignment of single-cell colony B bulk RNA-seq data from 5 in vitro time points projected onto the pseudo-time differentiation map. The alignment of bulk RNA-seq data is shown in the red line (right). e Expression analysis of differentially regulated genes in mouse CVSC single-cell colonies at different days after aggregation and culture