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Immunoprecipitation (IP) For immunoprecipitation, cells were lysed in HEPES-lysis buffer (50 mM HEPES, 150 mM NaCl, 1.5 mM MgCl 2 , 5 mM EDTA, 10% glycerol and 1% Triton X-100, pH7.4) supplemented with a protease inhibitor cocktail (Selleck Chemicals), and then centrifuged at 14,000 g for 15 min

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Time was a repeated effect with horse (treatment) as the subject