Fig. 3

Aging ltGLICOs are characterized by increased cellular stress and oxidative DNA damage. a Scaled cell stress signature scores in H1-derived ltGLICO cells grouped by cell type and sample age. b Quantification of total ROS levels measured by 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA)—assay. Graph shows mean + s.e.m. from four biological replicates. p-value = 0.0286, Mann–Whitney test. c Mitosox-Red staining for mitochondrial reactive oxygen species (ROS) levels in representative sections from young (4 month) and old (12 month) ltGLICOs; MitoSOX (red), and DAPI (Blue). d Immunofluorescence staining of ltGLICOs for 8-oxoDG (8-oxoDG in red, Phalloidin in green), representative images from a 2, 4, 9 and 18 month ltGLICOs. e Immunofluorescence staining of ltGLICOs for phospho-gamma-H2AX (yH2AX in red, DAPI in blue), representative images from a young (1.5 months) and old (18 month) ltGLICOs. Right, quantification of yH2AX positive cells per field of view at each CO age (plot shows mean + s.e.m.; 3 technical replicates). Scalebar = 200 µm