Figure 3.

HBO helps HCG suppress both bulk cancer cells and CSCs under hypoxic conditions. A) Relative cell viability of 4T1 cells treated with HCG under hypoxia or hypoxia + HBO conditions. B) Relative H₂O₂ content, ATP level, and C) ROS level of HCG‐treated 4T1 cells incubated in hypoxia or hypoxia + HBO conditions. D,E) Analysis of apoptosis in 4T1 cells after different treatments by flow cytometry. F) CLSM images of 4T1 cells after different treatments and stained with pH fluorescent probe BCECF. Scale bars: 50 µm. G) CLSM images (scale bars: 50 µm) and H) flow cytometry analysis of DOX fluorescence in 4T1 cells after treated with HCG under hypoxia or hypoxia + HBO conditions. I) Relative viability of CSCs treated with HCG in hypoxia or hypoxia + HBO conditions. J) The spheroid growth of 4T1 cells seeded in soft 3D fibrin with different treatments. Scale bars: 20 µm. K) ROS level in CSCs after different treatments. L) The colony number and M) colony size at day 7. Statistical significance was calculated by t‐test. p values: * p < 0.05, ** p < 0.01, *** p < 0.001, ns stands for not significant.