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Collection of in vivo peritoneal cytokines described as above. Serum cytokines were measured from blood collected via retro-orbital bleeds from indicated animals and centrifuged to separate the serum fraction. For in vitro cytokine secretion experiments, CAR T cells were cocultured 1:1 with ID8-Muc16ecto tumor cells (E:T ratio). Supernatant was collected after 16 hr of coculture and frozen at −80 °C until analysis. Cytokine detection was performed using the MILLIPLEX MAP Mouse Cytokine/Chemokine, Premixed 13 Plex kit, and the Luminex IS100 system.
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