Fig. 3

Cytotoxicity of Pep42-BBZ CAR-T cells towards GSCs in vitro. A Immunofluorescence (IF) staining of U-251MG CSC, GSC3# and GSC12# for csGRP78. Thirty-six hours prior to IF, cells were adhered to laminin-coated coverslips. The staining procedure was the same as for U-251MG cells. Scale bars = 10 μm. B Quantification of the relative cytotoxicity of CAR-T cells to the three GSCs lines. Target cells were cultured on laminin-coated coverslips overnight before co-culturing. CAR-T cells were co-cultured with target cells for 20 h at increasing E:T ratios of 1:1, 2:1, 4:1, and 8:1. All effector cells used in the assay were based on total T cells in each group. The cytotoxicity ratio was calculated based on flow cytometry results as described in the "Materials and methods" section. C ELISA-mediated quantification of IFN-γ release levels in the by supernatants from cells co-cultured at an E:T ratio of 4:1. Data presented as the mean volume ± SD, ** P < 0.01 *** P < 0.001