Muse® Cell Cycle Assay Kit

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	- Cells were pretreated with 100 or 200 µg/ml of EaEAC for 2 h and followed by 1 µg/ml of LPS for 24 h. - Cells were fixed with 70% ethanol at −20°C for 3 h

Protocol tips
- Cells were pretreated with 100 or 200 µg/ml of EaEAC for 2 h and followed by 1 µg/ml of LPS for 24 h. - Cells were fixed with 70% ethanol at −20°C for 3 h

Publication protocol

The cell cycle was measured using a Muse™ Cell Cycle kit (cat. no. MCH100106; EMD Millipore, Billerica, MA, USA) according to the manufacturer's instructions. Briefly, RAW264.7 cells were divided into 100 mm2 dishes (2.5×106 cells/dish), then pretreated with 100 or 200 µg/ml of EaEAC for 2 h. Following treatment with 1 µg/ml of LPS for 24 h, cells from subset groups were harvested by centrifugation at 3,000 × g for 5 min, then fixed with 70% ethanol at −20°C for 3 h. The fixed cells were washed with 1X PBS, then added to 200 µl Cell Cycle Reagent. Following incubation at 37°C in a CO2 incubator for 30 min, cell cycles were analyzed using a Muse® Cell Analyzer with the Muse 1.3.1 analysis program (EMD Millipore).

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