Serial analyses to confirm the mouse lines in the study. Presented is an example analysis performed for transgenic line ‘A’, carrying the human SNCA-Rep1 261 bp allele. (A) FISH analysis of metaphase chromosomes obtained from the mouse's spleen was used to determine the insertion site and to estimate the range of the copy number. Red-labeled human-PAC 27M07; green-labeled mouse BAC corresponding to mouse chromosome 9, used as a reference for copy number estimation; blue-DAPI. (B) RT–PCR to the 27M07 PAC was used to confirm the RNA expression of the human SNCA in the brain of an F1 mouse. (C) Western blot using the mouse monoclonal LB509 anti-human-SNCA (Zymed), to confirm the protein expression of the human SNCA in the brain of an F1 mouse. For (B) and (C), each lane represents analysis performed using a mouse-brain sample harvested from the following mouse lines: Rep1 261-A, the 261/261 line ‘A’ transgenic; PAC, control PAC containing the original Rep1 allele transgenic (55); Wt, wild type FVB; hSNCA, recombinant human SNCA protein.