Skip to main content
. 2022 May 5;29(5):826–839.e9. doi: 10.1016/j.stem.2022.04.001

Figure 3.

Figure 3

Lineage tracing and scRNA-seq by Gif+ chief cells identify injury-responsive chief cells

(A) Diagram of the Gif-Cre-nTnG allele used in this study.

(B) Experimental scheme of the L635 treatment study in Gif-Cre-nTnG mice. Dox was administered for 1 week and off for 2 weeks to label chief cells with GFP. The mice were then left untreated or treated with 3 doses of L635.

(C and D) Paraffin sections of Gif-Cre-nTnG mouse stomachs untreated or treated with 3 doses of L635 were immunolabeled with antibodies against (C) GFP (green), Mist1 (chief cells, red), GSII-lectin (neck cells, blue), and P120 (epithelial cell membrane, white) or (D) GFP (green), p57 (red), and Ki67 (proliferating cells, white). GFP+Mist1+ cells indicate chief cells and GFP+GSII+ cells indicate injury-responsive chief cells. Arrowheads indicate GFP+p57+ double-positive cells (yellow) and GFP+Ki67+p57-proliferating injury-responsive chief cells (red), respectively. Nuclei were counterstained with hoechst (blue) in (D). Yellow boxes indicate enlarged areas. Orange-dotted boxes indicate enlarged areas shown in Figure S3C. Scale bars, 100 μm.

(E) UMAP plots of scRNA-seq from Gif lineage cells (GFP+TdTom−) of Gif-Cre-nTnG mice in both conditions (Total, left), uninjured (Control, middle), and 2 days after DMP-777-induced corpus injury (Injury, right). Gifhigh and Giflow populations are marked by brown-dotted lines.

(F) UMAP plots for control (light blue) and injury (red) cells.

(G) UMAP plots of Gif expression in control (left) and injury (right).

(H) Dot plot for expression of marker genes of each cell type in Gifhigh and Giflow cell populations.

(I) Dot plot for expression of marker genes of proliferation or cell cycle in Gifhigh and Giflow cell populations.

(J) Projection of Gifhigh and Giflow cells on the UMAP plot for Pgc+ scRNA-seq. The cells in control and injury are denoted as light blue and red, respectively. Brown-dotted circles indicate the injury-responsive chief cell population.

(K) UMAP plots of p57 and Ki67 expressions in control (left) and injury (right).

See also Figure S3.