H22 and 4T1 cells were seeded in a 12-well plate at a density of 1×105 cells/well overnight under normoxic (21% O2) or hypoxic conditions overnight. For hypoxia groups, cells were cultured in a tri-gas incubator (Hua Xi Electronics Technetronic Co., Ltd, China) with a 2% oxygen concentration. The cells were then treated with Mn-TCPP, MOF or Mn-MOF at Zr concentration of 10 μg/mL and Mn concentration of 2 μg/mL for 10 h under normoxia or hypoxia. The cells were washed with PBS and then incubated with 5 μM 2'7'-DCFH-DA in dark for 30 min. The cells were irradiated with or without US (1 MHz, 0.9 W/cm2, 30% duty cycle) for 10 min under normoxia or hypoxia. After irradiation, the cells were washed with PBS three times and the intracellular ROS generation was determined by flow cytometry (FC500, Beckman Coulter, Fullerton, CA, USA) and Olympus FV1000 confocal microscope.

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