Fig. 3

A53T SynGFP inclusions co-localize with classical hallmarks of human Lewy pathology. a–g Mature A53T SynGFP inclusions are phosphorylated at serine-129 (pSyn), ubiquitinated, in an amyloid dye-binding configuration (X-34), co-stain with an anti-GFP antibody, and a small subset stain positively for cell death markers (Casp3, TUNEL). There was also a small subset of inclusions that were A53T SynGFP positive but did not stain for pSyn (e), and often these inclusions show signs of undergoing cell death processes as evidenced by their small, condensed nuclei (DAPI in e). Scale bar 5 µm. h–k A53T SynGFP inclusions also share the characteristics of Lewy pathology found in human genetic cases with the A53T mutation. Tissue from PFF-injected A53T mouse samples and human clinical samples contain somatic (arrow) and neuritic (arrow head) inclusions that stain with alpha-synuclein antibodies that selectively detect pathologic synuclein (Syn303 and Syn505), are ubiquitinated, pSyn positive, and increased in number compared to both mouse wild-type and human control PD-patient tissue. Scale bar 50 µm