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The medium was discarded from the petri dish, and the SH-SY5Y cells were rinsed with 500 µL PBS solution. Then, 250 µL of mixed lysate was added to each dish, and the SH-SY5Y cells were lysed for 0.5 h. Centrifugation was performed at 12,000 rpm/min for 10 min to collect the supernatant. The protein concentration of the samples was detected by using a Nano Drop System.
The following operations were the same as “Western blot of brain in mice”.
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