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. 2020 Aug 13;14:817. doi: 10.3389/fnins.2020.00817

FIGURE 2.

FIGURE 2

Glial cells were activated, which resulted in loss of motor neurons. (A,B) Western blot analysis was employed to detect the protein levels of GFAP, Iba1, and NeuN at the ages of 60 days (in the presymptom stage of ALS), 85 days (in the onset stage of ALS), and 117 days (in the end stage of ALS). The optical densities of GFAP, Iba1, and NeuN were analyzed with ImageJ software. (C,E,G) IHC was used to detect the morphology of astrocytes, microglia, and neurons in the lumbar anterior horns of SOD1G93A mice at different stages. (I) Positive staining for GFAP, Iba1, and NeuN in the anterior horn of the spinal cord was statistically analyzed with Image-Pro Plus (IPP). (D,F,H) Immunofluorescence was used to detect morphological changes in astrocytes, microglia, and neurons via immunostaining for GFAP, Iba1, and NeuN. (J) The astrocytes, microglia, and neurons in the lumbar anterior horns of SOD1G93A mice at different stages were counted. The data represent the means ± SEs from independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001, compared with non-transgenic mice.