Fig. 5

Flow cytometry characterisation of cell death by apoptosis (Annexin V binding) and necrosis (Propidium Iodide marking). a Representative flow cytometry output. Stained untreated or shaded controls showed significantly less binding of Annexin V or marking with PI relative to treated cells for all insults and conditions. None of the insults induced a significant degree of primary necrosis, with some PI marking of late apoptotic cells particularly in cells treated with staurosporine for 24 h. A clear second population of Annexin V labelled apoptotic cells was evident in stressed cells, with the separation between the apoptotic and non-apoptotic populations becoming more pronounced in the higher glucose environment. b A high glucose environment was associated with significantly higher levels of binding for Annexin V (early apoptotic cells) and combined Annexin V/PI (ie total of apoptotic, necrotic and late apoptotic/necrotic cells) than low glucose conditions for all insults. * indicate P < 0.05 between cells treated in the presence of 5mM and 25mM glucose