Fig. 3: Different pattern of cell death between myoblasts and myotubes in the co-culture with CTLs.
From: Targeting necroptosis in muscle fibers ameliorates inflammatory myopathies
a, b The representative confocal z-stack images of time-lapse analysis of H2KbOVA-myoblasts (a) or H2KbOVA-myotubes (b) co-cultured with OT-I CTLs in the presence of Annexin V (green) and PI (red). Nuclei of the cells were stained with Hoechst 33342 (blue). Images were taken at the indicated times from the starting of the co-culture. The cells surrounded by dotted line indicate dying myoblast and myotube. Scale bars indicate 20 μm. Representative data of three independent experiments are shown. c, d The TUNEL staining (red) of H2KbOVA-myoblasts (c) or H2KbOVA-myotubes (d) co-cultured with OT-I CTLs for 4 or 16 h, respectively. The myoblasts and myotubes were pre-labelled with CellTracker (green). Confocal z-stack images are shown. Scale bars indicate 20 μm. Representative data of two independent experiments are shown. e The immunofluorescence staining of FAS, CFLAR, RIPK1, RIPK3, MLKL, and phosphorylated MLKL at S345 (phospho-S345 MLKL; green) in H2KbOVA-myotubes co-cultured with OT-I CTLs for 8 h. Nuclei were counterstained with DAPI (blue). Scale bar indicates 10 μm. Representative data of three independent experiments are shown.
