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On the next day, after the membrane was washed with TBST for five times, HRP-conjugated secondary antibody (Thermo Scientific) was applied for 1.5 h at room temperature

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P3 Nucleofector Solution (Lonza, PBP3-00675) was prepared by mixing 16.4 μl of P3 Solution with 3.6 μl of Supplement 1 and the solution was allowed to equilibrate to room temperature