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For ectopic expression of proteins in mammalian cells, genes were inserted into pEGFPC1 (Clontech) or p4xFlagCMV 6

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Moreover, human glioma cells often express immune escape sites on the surface, such as PD-L1 and B7 homologue 3 (B7-H3), which can bind to immune checkpoint receptors, thereby inhibiting the activity and function of T cells (182)

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A second polymerase-based amplification reaction using a second set of primers, F\ and R 2 , is used to generate a blunt-ended product (labeled Reaction 2, Product 2), which is essentially identical to Product B