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For SDS-gel electrophoresis proteins were isolated from four to eight cervical spinal cord samples (p40) per genotype using Cell lysis buffer (#9803S, Cell Signaling Technology, Danvers, Massachusetts, USA) supplemented with protease inhibitor (#11697498001, Merck, Darmstadt, Germany)

The membrane was blocked with PBS containing 0.5% tween (PBST) and 5% milk, and probed overnight with affinity-purified anti-GspD or anti-PilQ in PBST plus 3% BSA

Nevertheless, at high concentrations, and in the presence of catalytic metal ions such as ferric iron (Fe 3+ ), VC can also have a pro-oxidative behavior through an indirect contribution in the Fenton reaction [6]

treatment is dependent on personal preference and experience of the treating doctor