Fig. 2
From: Reactive astrocytes undergo M1 microglia/macrohpages-induced necroptosis in spinal cord injury
Necroptosis of reactive astrocytes in injured mice spinal cord. a Western-blotting of RIP3 after SCI. Notice that the increase of RIP3 from 3 to 7 days post-injury. *P <0.05, **P <0.01. n = 3. b Double staining and quantification of RIP3 and GFAP at 5 days post-injury. Notice that at all time points, most RIP3-positive cells were GFAP-positive. Arrows points to RIP3/GFAP double-positive cells. Bar = 100 μm. Bar in insert = 50 μm. c Double-staining and quantification of RIP3 with OX42, NeuN and CC1. Notice that micoglia, neurons and oligodendrocytes account for a small portion of RIP3-positive cells. Bars = 50 μm. d, e Double staining of GFAP with MLKL and HMGB1. Many astrocytes around lesion center express MLKL and cytoplasmic HMGB1, shown by arrows. Bars = 100 μm. Bars in inset = 50 μm. f Immuno-eletron microscopic study of RIP3. Notice the focal cytoplasmic lysis of astrocyte (asterisks) and localization of RIP3 immunoreactivity on glial fibrils (arrows). Bar = 1 μm. g Immuno-eletron microscopic study of MLKL. Notice the fibrils (arrows in G1) and membrane localization (arrows in G2) of MLKL. Focal cytoplasmic lysis was shown by asterisks. Bar = 2 μm