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Closed recirculation system (not-evaporative) In a closed recirculation system, water is recirculated in a close circuit with negligible evaporation or exposure to the atmosphere or other influences that would affect the chemistry of the water in the system

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Western blotting Cells were lysed using RIPA buffer (Thermo Fisher Scientific, Waltham, MA, USA) containing protease inhibitors and phosphatase inhibitors (Beyotime, Nanjing, China), and protein concentrations were measured using a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA)