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Cells were incubated with serum-free media for 24 h prior to each experiment and seeded at 1 10 6 cells/dish in 100 mm culture dishes for western blotting and 3 10 4 cells/well in 24-well- culture plates for immunofluorescence

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Then, the samples were frozen immediately and stored at 80C

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do not store pens with needles attached, as this can cause medication leakage or air bubble formation