Gentamycin sulfate

Dulbecco’s PBS(−), heart-infusion broth, and trypticase soy broth were purchased from Nissui Pharmaceutical (Tokyo, Japan). HBSS supplemented with Ca²⁺ and Mg²⁺ was purchased from Gibco Invitrogen (Life Technologies, Carlsbad, CA, USA). Gentamycin sulfate, trisodium citrate dihydrate, dextran (MW ∼200,000 Da), paraformaldehyde, glutaraldehyde, OsO₄, and RIPA buffer containing a protease inhibitor cocktail were purchased from Nacalai Tesque (Kyoto, Japan). Cytochrome c, apocynin, DPI, sheep serum, and zymosan were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bafilomycin A1, glycine, SDS, 2-amino-2-hydroxymethyl-1,3-propanediol [Tris (hydroxymethyl) aminomethane], SOD, and polyoxyethylene [10 (link)] octylphenyl ether (Triton X-100) were purchased from Wako Pure Chemical Industries (Osaka, Japan). DNase was purchased from Roche (Basel, Switzerland).

As previously described [36 (link)], mice were treated with antibiotics, which were administered daily starting from seven days before the injection of α-synuclein PFF until the end of the experiments. The antibiotics were administered as a cocktail provided in sterile drinking water and included ampicillin (1 g/L; Nacalai Tesque, Kyoto, Japan), vancomycin (0.5 g/L; Shionogi Pharma), neomycin (0.5 g/L; Nacalai Tesque), gentamycin sulfate (100 mg/L; Nacalai Tesque) and erythromycin (10 mg/L; Sigma-Aldrich Japan, Tokyo, Japan). We prepared stock solutions of ampicillin, vancomycin and neomycin by dissolving them in sterilized water at a concentration of 50 mg/mL. erythromycin was dissolved in 99.5% ethanol (FUJIFILM Wako Pure Chemical) at a concentration of 50 mg/mL, while gentamycin was dissolved in sterilized water at concentration of 25 mg/mL. The stock solutions were aliquoted and stored at −30 °C for two months. For preparation of an antibiotic cocktail, the stock solutions were mixed with 300 mL of sterilized water. The average amount of water intake was 5 to 7 mL/mouse/day.