Fig. 5: CAR T killing of HeLa target cells is mediated by soluble factors including TNFα.
From: Quantifying requirements for mitochondrial apoptosis in CAR T killing of cancer cells
A, B Conditioned media was collected as described in the text and incubated with HeLa-19 or HeLa-DKO-19 target cells for 24 h. For A, HeLa N = 6, DKO N = 3; in B HeLa N = 4, DKO N = 1. In C, conditioned media from two paired co-cultures (HeLa-19 or HeLa-DKO-19 with 28z CAR T cells) was incubated with HeLa-19 or HeLa-DKO-19 target cells for 24 h. N = 1. Figures D and E present Cytokine Array data from conditioned media with intensity normalized to the included positive and negative controls. Figure D highlights cytokines that were unique to HeLa-19 co-culture with a specific CAR by subtracting the cytokines present in HeLa-19/Δζ co-culture (N = 1). Figure E presents results from all three cytokine arrays performed, highlighting the top cytokines noted in Figure D. F Annexin V / Hoechst viability assay performed 24 h after HeLa-19 incubation with conditioned media and the indicated blocking antibodies, N = 3. Each point represents a biological replicate, significance corresponds to unpaired t test, **p = 0.0017.
