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5.8 Flow cytometry To verify intracellular expression of antigens by the pDNA3.1 plasmid human embryonic kidney cells (293T) were cultured in six-well plates in RPMI-1640 medium (R0883-500ML, Sigma-Aldrich) supplemented with 10% heat-inactivated FCS, 2mM L-glutamine, and 100 U/ml penicillin-streptomycin at 37C until they reached 70-80% confluency