"Designing the Optimal Artificial Immune Synapse: Combining TCEs with Diakines" Image Files

Internalization Assay Human CD8⁺ T cells incubated with Dylight650-labeled DK210 (EGFR) along with FM™ 1-43 Dye (N-(3-Triethylammoniumpropyl)-4-(4-(Dibutylamino) Styryl) Pyridinium Dibromide). For conditions with neutralizing antibodies, anti-IL2Ra (R&D Systems) and anti-IL-10R1 (BioLegend) were incubated prior to DK210 (EGFR) addition. Synapse Imaging Human CD8+ T cells were cultured with Dylight650-labeled DK210 (EGFR) then cultured with A498-GFP and CD3xEGFR T-cell engager. Human CD8+ T cells were labeled with MemBrite580 (Biotium). TCR Co-localization Human CD8+ T cells were cultured with DylightTM 550-conjugated DK210 (EGFR) (100 ng/mL). T cells were stained with APC conjugated anti-CD3 prior to coculture with GFP Raji-B cells. Instrument: Images were taken using a Yokogawa CSU X-1 spinning disc confocal microscope.