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After incubation with the secondary antibody, the cells were washed with PBST and mounted using a Fluoroshield Mounting Medium with DAPI (Abcam ab104139)

The following sections break down each scenario

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Cells were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS, L-glutamine, penicillin, and streptomycin at 37 C in 5% CO 2