Adult animals were washed three times in 1.5 ml S-basal buffer and 50 200 animals were placed with glass Pasteur pipette near the center of an NGM plate, equidistant from the two bacteria.
To substantiate whether differences in activation of coagulation, downregulation of fibrinolysis, viral loads and TNF-α and IL-12p70 levels between rm-APC and buffer control treated animals were associated with an altered mortality we performed a survival study.
After thirty minutes, six were treated with 0.5 mg IdeS in PBS, but this time the enzyme was injected i.v.The remaining six mice were given PBS alone, also i.v. Again the buffer treated animals were all dead within 24 hours, whereas the IdeS treated animals remained healthy and unaffected until sacrificed two weeks later.
One week after the STZ or citrate buffer injection, animals were exercised.
One week after the STZ or citrate buffer injection the animals were exercised.
After removing the odorant by washing the worms with S-basal buffer, animals were placed on an assay plate, with an ethanol-diluted butanone point source opposite from an ethanol point source, and were allowed to roam for 2 h at 20°C.
At one week after the initial injections of STZ or citrate buffer, animals were then treated with intraperitoneal injections of either saline, PEG-SOD (2,000 units/kg/day), or denatured PEG-SOD (heat denatured 2,000 units/kg/day) for a total of seven days.
To maintain the phosphorylated activity, all phosphate buffers were avoided, and instead animals were washed (five times) in Tris-buffered saline with 0.1%Tween200 (TBST buffer).
After dissection of the abdomen, legs, and proboscis in 4% glutardialdehyde in 0.1 M cacodylate buffer at 4°C, the animals were fixed in 4% glutardialdehyde and 1% tannin in 0.1 M cacodylate buffer at 4°C and stored under refrigeration at 4°C.
