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Pooled cells in vials were thawed, counted and seeded into wells of six-well plates in RPMI-1640 without phenol red (Gibco, 11835030) supplemented with 10% FBS (Sigma-Aldrich, F7524) at a seeding density of 375,000 cells per well

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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