Fig 1: iRGD-anti-CD3 promoted T-cell activation and cytotoxicity. (A) Flow cytometry results and (B) quantification of CD69 and CD25 expression on CD8+ and CD4+ T-cell subsets. (C) Assessment of IFN-? and TNF-a in coculture supernatants. (D) Lysis of monolayer tumor cells at indicated E:T ratios. (E) Confocal microscopy images (upper panel) and surface display (lower panel) of live/dead viability assays in MCSs. Live cells were stained with calcein acetoxymethyl ester (AM) (green) and dead cells with EthD-1 (red); magnification, ×200; scale bar, 50 µm. (F) Live/dead cell quantification in MCSs. Data are represented as mean±SEM; n=5. Student’s t-test; **p<0.01, ***p<0.001, ****p<0.0001. CG7C-, CG7C-anti-CD3; RGD-, RGD-anti-CD3; iRGD-, iRGD-anti-CD3. E:T, effector-to-target; IFN-?, interferon gamma; MCS, multicellular spheroid; TNF-a, tumor necrosis factor alpha.