Fig. 6

In vitro cytotoxicity and ferroptosis effect of MSF@CCM. a Schematic illustration of MSF@CCM-enhanced cytotoxicity and ferroptosis via producing ROS and LPO. b Cell viability of NIH-3T3 cells treated with MSF and MSF@CCM at different concentrations. Data are presented as mean ± SD (n = 4). c Cell viability of 4T1 cells treated with MSF and MSF@CCM at different concentrations with or without US irradiation. US: 1.0 MHz, 1.5 W cm⁻², 2 min, 50% duty cycle. Data are presented as mean ± SD (n = 4). d CLSM images of 4T1 cells co-stained by Calcein AM/PI. Green (Calcein AM), live cells; red (PI), dead cells. US: 1.0 MHz, 1.5 W cm⁻², 2 min, 50% duty cycle. Scale bar = 100 μm. e CLSM images of 4T1 cells incubated with FITC-labeled MSF and MSF@CCM for different time points. Scale bar = 25 μm. f Corresponding mean green fluorescence intensity of MSF and MSF@CCM in (e). Data are presented as mean ± SD (n = 3). g CLSM images and (h) corresponding fluorescence intensity of ROS with different treatments. Green (DCFH-DA), ROS. US: 1.0 MHz, 1.5 W cm⁻², 2 min, 50% duty cycle. Scale bar = 100 μm. Data are presented as mean ± SD (n = 3). i Bio-TEM observation of mitochondrial morphology of 4T1 tumor cells after different treatment. Scale bar = 1 μm. Scale bar for localized magnification is 200 nm. j Intracellular GSH levels in 4T1 cells with different treatments. Data are presented as mean ± SD (n = 3). k CLSM images of cells stained by Liperfluo for monitoring intracellular LPO levels induced by different treatments. Scale bar = 50 μm. l Levels of MDA after different treatments. Data are presented as mean ± SD (n = 3). US: 1.0 MHz, 1.5 W cm⁻², 2 min, 50% duty cycle. One-way ANOVA with Tukey’s multiple comparisons was used to calculate statistical differences in (c, f, h, j, and l)