C3 tg prnp snca a53t 83vle j

Ethical treatment of animals followed AALAC and NIH guidelines on TTUHSC Institutional Animal Care and Use Committee (IACUC) protocol #12001. A53T aSyn wild type (WT) and transgenic (Tg) mice (B6;C3-Tg-Prnp/SNCA*A53T/83Vle/J; Jackson Laboratories, Bar Harbor, ME) were evaluated at 10 mo and 17 mo after receiving 5–12 mo of oral FTY720 (0.5 mg/kg/mouse) prepared in 200 proof EtOH (vehicle) given 2×/week, while control mice received vehicle alone on the same schedule (N = 17). WT and Tg CNP-aSyn MSA mice (B6;C3-Tg-CNP-SNCA-M2Vle) [25 (link)] were assessed at 10–11 mo after receiving 2 or 3 mo of FTY720-Mitoxy (1.1 mg/kg/day) prepared in 200 proof EtOH plus Lactated Ringers (vehicle) or vehicle alone, delivered by Alzet pump Model 2006, implanted subcutaneously on the back of the animal without interfering with normal movement of the mice (N = 10).

Anti-rabbit Bpoz-2 antibody (WB, IHC and IP; cat # ab112610), anti-rabbit αsyn antibody (cat # ab32518) and anti-PINK1 antibody (WB and IP: cat # ab75487) were purchased from Abcam. Anti-sheep α-syn antibody (Cat# AB5336P) and Rabbit-anti Tyrosine Hydroxylase (TH) antibody (Cat # AB152) were purchased from EMD Millipore. Bpoz-2 overexpression construct (cat # EX-Mm12657-Lv104), Bpoz-2-siRNA (cat# MSH042485-LvH1), lentiviral packaging kit, HEK-293Ta cells (Cat # CLv-PK-01), and chemically competent E-Coli cells (Cat # STK200-10) were purchased from Genecopoeia. C1A53T-tg animals (strain # B6;C3-Tg(Prnp-SNCA*A53T)83Vle/J) were purchased from Jackson Laboratory. These mice were genotyped and maintained in the animal care facility of Rush University Medical Center up to 8–9 moths of their age before viral injection. Animal experiments and viral injections had been performed in accordance with the guidelines and regulations imposed by Institutional Animal Care and Use Committee (IACUC) and Institutional Biosafety Committee (IBC) respectively. Experiments were performed after receiving necessary approvals from IACUC (protocol # 14-028) and IBC (protocol # 040814).