Fig. 3

The regulatory effects of iMSCs on oxidative stress levels and the expression of antioxidant genes and proteins in macrophages stimulated with LPS + IFN-γ. A ROS levels in macrophages were visualized using dichlorofluorescin diacetate staining, with bright-field and fluorescence microscopy imaging. B Flow cytometry analysis of fluorescence intensity in the FITC channel was performed to assess ROS levels across different treatment groups, with the left panel showing gating strategy and the right panel displaying fluorescence intensity distributions for each group. C Quantification of ROS levels was assessed by measuring the mean fluorescence intensity (FITC-mean) using flow cytometry. D The relative expression levels of antioxidant genes (CAT, HO-1, NQO1) in macrophages were determined by RT-qPCR at 24 and 48 h. (E) Protein expression levels of Nrf2, HO-1, and NQO1 were analyzed via Western blot (Full-length blots are presented in Supplementary Fig. 2A-D). F Quantitative analysis of Nrf2, HO-1, and NQO1 protein levels was conducted relative to β-tubulin expression. *P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant (P > 0.05)