Fig 1.
Localization and characterization of NG2 cells in the stomach of adult rats.
(A-B) Localization of NG2 cells in the gastric mucosa. (A) Immunofluorescent images showing staining with both monoclonal NG2 (green) and polyclonal NG2 (red) antibodies and Hoechst (blue). (B) Magnified view of immunofluorescent images showing staining for NG2 (red) and Hoechst (blue). (C-D) Colocalization of CD34 (green) in NG2+ telocytes (red) of the adult stomach. Immunofluorescence staining for NG2 (red), Hoechst (blue) and CD34 (green) (a telocyte marker). (E-F) Co-expression of PDGFRα (green) in NG2+ telocytes (red) in the gastric mucosa. Immunofluorescence staining for NG2 (red) and PDGFRα (green) (another telocyte marker). Arrowheads show NG2+/PDGFRα+ telocytes whereas arrows point to NG2+/PDGFRα- telocytes. (G-H) Triple immunofluorescence staining for PDGFRα (green), CD34 (red) and NG2 (blue). Arrowheads indicate NG2+/CD34+/PDGFRα+ telocytes and arrows show NG2+/CD34+/PDGFRα- telocytes. Scale bars: 100 μm (A, C, E, G) and 20 μm (B, D, F, H).
Fig 2.
Identification of NG2+ telocytes in rat stomach using electron microscopy.
(A-C) Representative electron micrograph of NG2-labeled gastric telocytes in the adult rat. (B-C) Magnified views of NG2 immunoreactive cell (arrow) in panel (A). (C) Arrowheads show cellular processes, stained with the anti-NG2 antibody. Scale bars: 10 μm (A-B) and 1 μm (C).
Fig 3.
Expression of the fLuc reporter gene in NG2 cells of Tg rat stomach.
(A-C) Co-expression of the fLuc transgene was observed in almost all gastric NG2+ cells of Tg rats. Immunofluorescence images of stomach sections stained with anti-NG2 (red) and anti-luciferase (green) antibodies and Hoechst (blue). Scale bars: 100 μm (A) and 20 μm (B-C).
Fig 4.
Ex vivo bioluminescence imaging of rNG2-fLuc Tg rats.
(A-B) Bioluminescence images of several organs removed from young (A) and aged (B) Tg rats. The area indicated by red dashed lines includes the stomach and shows regions of interest (ROI). Upper panels: Photographs of several organs. Lower panels: Bioluminescence images of these organs. (C) Quantification of bioluminescence signals from the isolated stomach of young (light blue bar, n = 8) and aged (pink bar, n = 8) Tg rats.
Fig 5.
Comparison of NG2 gene expression in the stomach of young and aged Tg rats.
NG2 expression in young (light blue bar, n = 6) and aged (pink bar, n = 6) stomach of Tg rats. Data are mean ± SEM. * p < 0.05.
Fig 6.
Age-related changes in NG2+ telocytes in the rat stomach.
(A-F) Double immunofluorescence images of young (A, C, and E) and aged (B, D, and F) rat stomach sections stained with anti-NG2 (blue, A and B) and anti-PDGFRα (green, C and D) antibodies. (E, F) Merged images of NG2 (blue) and PDGFRα (green). Scale bar: 100 μm.