Figure 3. Identification of human stem cell sub-clusters by snRNA-seq.

(A) UMAP showing the stem cell cluster identification based on the snRNA-seq data from the six merged human pituitary samples. Each cell cluster is color-coded and numbered. GATA3 lineage-committed progenitor stem cells are circled.

(B) Feature plots depicting the expression distribution of key stem cell marker genes and of cell lineage commitment marker genes among the various clusters. A scale is included for each feature plot. All scales are similar except for POMC due to background gene expression. For the GATA3, POMC, and POU1F1 committing cell lineages, clusters are labeled for easier identification and localization of the cells throughout the PSC clusters. Additional gene feature plots are presented in Figure S3.

(C) UMAPs identifying all color-coded stem cell sub-clusters in females and males. The feature plot on the far right shows XIST expression, highlighting the female samples.

(D) UMAPs identifying all color-coded stem cell sub-clusters in the pediatric, adult, and aged donors.

(E) Colocalization of Sox2 (red) and Jun (blue) transcripts in a wild-type P56 CD-1 male adult mouse pituitary. Scale bar, 200 μm. AL, anterior lobe; IL, intermediate lobe; PP, posterior pituitary. Top, full image. Bottom, magnification of the boxed region on the top panel. Arrows highlight specific cells with colocalization of Sox2 and Jun. Refer to Figure S3 for Sox2 and Jun colocalization at P3 and P15. Shown is a representative staining from three to five biological replicates.

(F) Chromatin accessibility track analysis and gene expression analysis (violin plots to the right of the chromatin accessibility tracks) and for SOX2 and GATA3 in all pituitary cell types from the sn multiome dataset generated from the pediatric female. The gene structure is provided below the tracks.