Fig. 4

Synergistic ferrotherapy. (a) ^•OH staining by a BBoxiProbe®O26 fluorescent probe in 4T1 cells after different treatments. (b) Lipid peroxides imaging by a Liperfluo fluorescent probe. (c) Flow cytometry analysis of the LPO fluorescence intensity of 4T1 cells subjected to different treatments. (d) Measurement of intracellular MDA levels. (e, f) Images of cell comet electrophoresis assay (e) and percentage of fluorescence intensity (Tail DNA%) in the comet tail (f) in 4T1 cells after different treatments. (g) Western-blot analysis of the expression of γ-H2AX. (h) Flow cytometry analysis of the apoptosis of 4T1 cells after different treatments. (i) Quantitative analysis of cell cycle distribution by flow cytometer. (j, k) Western-blot analysis of the expression of proteins that related to ferroptosis (j) and ferritinophagy (k). (1), (2), (3), and (4) indicate the groups of control, ActD, FessMOF-PEG, and FessMOF/ActD-PEG, respectively. All data were presented as mean ± standard deviation. Statistical differences were calculated using two-tailed Student’s t test. Differences were considered significant when the p-value was less than or equal to 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001