Fig. 4: Morphology of megakaryocytes in response to TNF-induced necroptosis and apoptosis.
From: The necroptotic cell death pathway operates in megakaryocytes, but not in platelet synthesis
a Representative time-lapse confocal images from WT BM-derived cultured megakaryocytes treated with TNF (100 ng/ml, T), Smac mimetic (0.5 µM, S), QVD-OPh (50 µM, Q), Necrostatin-1 (50 µM, N), and ABT-737 (5 µM). Megakaryocytes were labelled with CD41 conjugated to Alexa-488 (green) and PI (propidium iodide, red), Bar 50 µm. Cells were maintained on the stage within a humidified chamber at 37 °C and 5% CO2 for 7 h. Images were acquired on a Ziess live cell Axio inverted modular microscope for live cell imaging every 10 min driven by AxioVision v4.8 software. b Representative time-lapse confocal images from WT and Mlkl−/− BM-derived cultured megakaryocytes treated with the indicated agents: TNF (100 ng/ml, T), Smac mimetic (0.5 µM, S), IDN-6556 (5 µM, I), Necrostatin-1 (50 µM, N) at the indicated time points (hours) at 37 °C, images acquired as in (a). c Quantification of PI from representative time-lapse video microscopy at the indicated time points (hours). PI fluorescence intensity was quantified using a custom macro in Fiji software. n = 3 independent experiments, data presented as mean ± SEM, One-way ANOVA with the Bonferroni multiple comparison test, **P < 0.005; ***P < 0.001.
