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. 2020 Aug 28;217(12):e20191166. doi: 10.1084/jem.20191166

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© 2020 Oda et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure S5. — IncuCyte assay. (A) NucLight Red⁺ Panc1 cells were cultured alone (white) or co-cultured with TCR-only (black), TCR/trFas (blue), or TCR/Fas-4-1BB (red) T cells at a 30:1 E:T ratio. Tumor cell concentration was determined by Total Red Area (µm²/image). Averaged data represent T cells generated from three unique human donors. Statistical analyses are shown: TCR/Fas_tm-4-1BB versus TCR only, *, P< 0.05; TCR/trFas_tm-4-1BB versus TCR only, P = not significant; t test of final time point. (B) Lamp-1 (CD107a) expression of T cells previously exposed to tumor. T cells previously stimulated in the IncuCyte assay (as in A) were restimulated with Panc1 cells for 5 h and then stained for surface and intracellular Lamp-1 (CD107a) before evaluation by flow cytometry. Averaged data represent T cells generated from three unique human donors. TCR only versus TCR/trFas: *, P< 0.05; TCR only versus TCR/Fas_tm-4-1BB: P= not significant; t test. gMFI, geometric MFI.