Fig. 5

Cuprizone induces diverse forms of cell death. a Schematic of acridine orange (AO) fluorescence for lytic and apoptotic cells from [97]. b Representative images from AO-stained tissue highlighting green nuclear (DNA) and red cytoplasmic (RNA) fluorescence. (c) Representative AO images of ‘early apoptosis’ and ‘late apoptosis’ morphologies based on red condensing and fragmented morphologies, respectively. AO images were matched with comparable immunohistograph images with the apoptotic marker cleaved caspase 3 (CC3). (d) Early apoptotic, (e) late apoptotic, and (f) lytic cells stratified by CC3 expression were quantified from control animals and those on a cuprizone diet. g Representative morphologies of early apoptotic, late apoptotic, and lytic cells based on AO fluorescence. h Quantification of cell death based on AO fluorescence across a five-week cuprizone time course. (d-f) n = 2–3 (h) n = 3–7; Two-way ANOVA with Tukey’s multiple comparisons test (p < 0.05). Error bar ± SEM. Scale bar, (b) 50 μm (c, g) 10 μm