Fig. 2: Loss of Bak and Bax confers resistance to CAR T killing of HeLa target cells.
From: Quantifying requirements for mitochondrial apoptosis in CAR T killing of cancer cells
A Annexin V / Hoechst viability staining of HeLa-19 and HeLa-DKO-19 cells following 48 hour exposure to etoposide, N = 3. B BH3 profiling assay comparing mitochondrial response to the Bim peptide in HeLa-19 (N = 2) and HeLa-DKO-19 (N = 1) cell lines. Figure C–F depicts target cell viability following exposure to CD19 CAR T cells assessed via annexin V/Hoechst staining at 20 h (C, D), an impedance assay (E, N = 2), and a colony formation assay (F, N = 1). C presents data from 4 biological replicates using a single donor’s CAR T cells. Curves were fitted with non-linear regression, error bars are technical replicates. All biological replicates are shown in Fig. S2. The left panel of D presents the E:T 50 value for every paired co-culture experiment performed (N = 20 for 28z, N = 12 for BBz, 9 donors total). The right panel presents the mean difference in E:T50 (DKO-HeLA, middle bar) with the 95% confidence interval. For all graphs except panel C, each point is a biological replicate. Significance corresponds to either an unpaired (A, E) or paired (D) t test, ****p < 0.0001, **p = 0.0015.
