Fig. 3

In vitro cellular uptake and antitumor mechanism of MLNPs. (a) CLSM images and fluorescence distribution profiles (Scale bar = 50 μm). (b) CLSM cross-section images of 5-layered cellular uptake of DiO-labeled MLNPs (green) after incubation for 5 h. Hepa1-6 cells were labeled with DAPI (blue) and Rhodamine phalloidin (red) (Scale bar = 50 μm). (c) Percentages of DiO-labeled MLNPs internalized by Hepa1-6 cells for 1, 3, and 5 h, respectively. Each point represents the mean ± S.E.M. (n = 3). (d) CLSM images and (f) quantification of ROS changes in Hepa1-6 cells labeled with Hoechst 33,342 (blue) after co-incubation with MLNPs for 6, 12, and 24 h, respectively (Scale bar = 50 μm). Each point represents the mean ± S.E.M. (n = 3). (e) CLSM images and (g) quantification of JC-1 and Hoechst 33,342 stained Hepa1-6 cells after co-incubation with MLNPs for 12, 24, and 48 h, respectively (Scale bar = 20 μm). Each point represents the mean ± S.E.M. (n = 4). (h) Cell cycle analysis of Hepa1-6 cells after co-incubation with MLNPs for 12 or 24 h by FCM. Each point represents the mean ± S.E.M. (n = 3; *p < 0.05). (i) Schematic illustration of the pro-apoptotic mechanism of MLNPs against liver tumor cells