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Complement lysis. All antisera were heated at 56°C for 30 min before use, and fresh RS served as the source of complement. Western blotting to detect the level of cytoplasmic protein PNP was performed to monitor the lysis of M. bovis following treatment with normal RS (NS), anti-P81 serum, anti-UgpB serum, or anti-M. bovis serum. M. bovis was incubated with RS at a 1:10 dilution as the source of complement or with HIRS for 1 h (A) or 3 h (B).
