Figure 4
UBQLN2 interacts with and may alter α-syn levels in A53T mice. (A) Schematic depicting breeding strategy to obtain A53TxUb2-hi and A53TxUb2-KO mice. (B) Western blot of RIPA-soluble brain lysates showing increased levels of total α-syn in A53TxUb2-KO mice (n = 6), while A53TxUb2-hi (n = 7) mice show a trend toward decreased α-syn levels compared to non-transgenic (Non-Tg) controls (n = 8). (C) Representative images of total a-syn (magenta) and UBQLN2 (cyan) immunofluorescence in the deep cerebellar nucleus of 12-month-old Non-Tg (n = 6), A53T (n = 6), A53TxUb2-hi (n = 6), and A53TxUb2-KO (n = 5) mice and quantification show no significant difference in total a-syn expression between genotypes. (D) Western blot showing total endogenous α-syn levels were unchanged in both Ub2-hi (n = 4) and Ub2-KO mice (n = 4) compared to Non-Tg controls (n = 4). Western blots were cropped to focus on protein(s) of interest. Uncropped Western blots are shown in Supplemental Figure S3). (E) Immunoblot showing endogenous and transgenic UBQLN2 is pulled down when α-syn is immunoprecipitated from mouse brain lysate (n = 4). (F) Immunoblot showing α-syn coprecipitates with FLAG-tagged UBQLN2 in A53TxUb2-hi mice when FLAG is immunoprecipitated from mouse brain lysate (n = 2).
