Fig. 2

In vitro anti-tumor effects of TLNTs. A Cytotoxicity of TLNTs against various tumor cell lines after co-incubation with TLNTs at protein concentrations from 0.5 to 64 µg/mL for 24 and 48 h, respectively. Each point represents the mean ± S.E.M. (n = 5). B Pro-apoptotic properties of TLNTs after co-incubation with TLNTs for 4 and 8 h, respectively. C CLSM images of 4T1 cells stained with DCFH-DA after co-incubation with TLNTs for 4 and 8 h, respectively (scale bar: 50 μm). D ROS fluorescence intensity of 4T1 cells after co-incubation with TLNTs for 4 and 8 h, respectively. E Mitochondrial membrane potential changes in 4T1 cells (scale bar: 50 μm). F TLNTs restrained cell cycle progression in 4T1 cells after co-incubation with TLNTs for 12 and 24 h, respectively. Populations of 4T1 cells in various cell cycle phases were determined by FCM. Each point represents the mean ± S.E.M. (n = 3; *p < 0.05 and **p < 0.01). G Western blot analysis of 4T1 cells receiving the treatment of TLNTs for 48 h. Cyclin A, cyclin B and cyclin D proteins were probed. GAPDH was probed to ensure the equal loading of total proteins in each lane