Fig. 4

Filamentous and granular α-synuclein inclusions form in MSA-infected cultured TgM83 astrocytes. 3D volume visualization of maximum intensity projection of Z-stacks of α-synuclein inclusions in MSA-infected TgM83^+/+ astrocytes aggregating as a filamentous and b granular α-synuclein inclusions. c, e Quantification of pSyn (S129) inclusions formed in MSA-infected TgM83^+/+ cultures [size I (< 10 μm), size II (10–50 μm), and size III (> 50 μm)] at 0, 7, 14, and 21 days post-exposure (dpe). Cell count of each group is represented as percentage of total cells, and the data were acquired from 16 randomized fields from four replicate experiments (n = 4). Astrocytes were immunostained for total human α-synuclein (αSyn, green), phosphorylated α-synuclein [pSyn (S129), red], and GFAP (white). d, f Maximum projection intensity of Z-stack and merge of all three channels is shown. Co-localization of pSyn (S129) and αSyn appears yellow. Nuclei were stained with DAPI (blue). Scale bars, a, b 20 μm, d 5 μm, f 10 μm