Fig. 3

MSLN-CAR T cells efficiently kill cancer stem-like cells. A, B Quantification of RNA transcript levels for CD133, CD44, CD117 and CD24 in ovarian cells (SKOV3, OVCAR3) and their cancer stem cell derivations (SKOV3-CSC and OVCAR3-CSC). For statistical analysis of two groups an unpaired two sample t-test is used. Data were plotted and are shown as mean ± SD; * P < 0.05, *** P < 0.001. C Quantification of RNA transcripts for MUC16 in ovarian cells (SKOV3, OVCAR3) and their cancer stem cell derivations (SKOV3-CSC and OVCAR3-CSC) were determined by confocal imaging. D, E Both flow cytometry and immunofluorescence staining were used to stain live cells. A–D consist of three technical replicates. The pictures represent one example of three technical replicates. F The cytotoxicity of MSLN-CAR T cells on luciferase-expressing target cells was quantified after incubation for 20 h at an E:T ratio of 2:1. Three independent experiments were performed per condition. G ELISAs were used to detect IFN-γ release by T cells in coculture supernatants. E and F consist of three biologic replicates and three technical replicates. Statistics: two-tailed one-way ANOVA. Data are presented as the mean ± SD, n = 3; ***P < 0.001 vs CD19-CAR or NTD