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. Author manuscript; available in PMC: 2019 Sep 3.
Published in final edited form as: J Vis Exp. 2019 Feb 27;(144):10.3791/59275. doi: 10.3791/59275

Figure 1. Schema and analysis strategy of repetitive challenge assay.

Figure 1.

(A) Schema and timeline of repetitive tumor challenge assay. For each well, CAR T cells were first co-cultured with PBT030–2 GBM cells (4,000 CAR+ cells, 16,000 tumor cells) and re-challenged with 32,000 tumor cells every other day (D2, D4 and D6). Analysis of tumor cell and CAR T cell number, as well as CAR T cell phenotype is carried out at D1, D3, D5 and D7. (B) Gating strategy to determine CAR% in T cells before setting up the co-culture. (C) Gating strategy of live cells, tumor cells and CAR T cells from repetitive challenge assay. (D) Tumor cells number at different times of the rechallenge assay, co-cultured with untransduced T cells. Error bars: ±SEM