Fig. 2

DARPin28z induces murine CAR-T cell cytokine production upon HER2 stimulation. a 10⁶ scFv28z, DARPin28z, or CAR –’ve transduced murine T cells were stimulated with HER2 (HER2Fc fusion protein) or an unrelated target (KDRFc fusion protein) for four hours at 37 °C in a 96-well plate. Production of IFN-γ and TNF-α was measured by intracellular cytokine staining (ICS) and subsequent flow cytometry. Data from CD8⁺ Thy1.1⁺ T cells is presented as mean of n = 3 experiments (CAR –’ve) and n = 5 experiments (scFv28z and DARPin28z) ± SD. Bracketed numbers are quantitative of the representative plots shown. b Visual comparison of cytokine production data from a. c Cytokine expression data expressed relative to CAR-positivity of transduced cells where CAR⁺ transformant = % CAR⁺/% Thy1.1⁺. Error bars = SD. * = p < 0.05 ** = p < 0.005 *** = p < 0.001