Fig. 2

Anti-TNBC efficacy of Nano-DOX in comparison with DOX. a Effects of Nano-DOX and DOX on the viability of 4T1 cells in vitro assayed by the CCK-8 test. b Effects of Nano-DOX and DOX on the proliferation of 4T1 cells in vitro assayed by CFSE staining. c, d Apoptosis of 4T1 cells after 24-h treatment of Nano-DOX or DOX assayed by annexin V immunofluorescent staining and FACS. e, f Size and weight of orthotopic 4T1 tumor xenografts in mice at the end of 3-week treatment of Nano-DOX or DOX. g Immunohistochemical staining of Ki67, PCNA (markers of cancer cell proliferation), and caspase 3 (marker of cancer cell apoptosis) in mouse orthotopic 4T1 tumor xenografts at the end of 3-week treatment of Nano-DOX or DOX. (Duration of Nano-DOX or DOX treatment was 24 h for the in vitro cell experiments.) In FACS analysis, geometric means were used to quantify fluorescence intensity. Values were mean ± SD (n = 3 for in vitro experiments and n = 8 for in vivo experiments, *p < 0.05, **p < 0.01)