Figure 5.

HO-1/BMMSC-mediated inhibition of ferroptosis and alleviation of SHP-HR injury could be mediated by regulating the protein levels of IREB2. Evaluation of ferroptosis and cell injury in SHP-HR with BMMSCs or HO-1/BMMSCs. (a) Representative immunofluorescence images of IREB2 showing that positive expression increased significantly in HR and was decreased by HO-1/BMMSC treatment. (b) IREB2/FTH1/TFR1/GPX4 protein levels assessed using western blotting. (c) Ireb2 mRNA level detected using qRT-PCR. (d) Fe²⁺ level detected using colorimetry. (e) The lipid ROS level detected using C11-BODIPY and flow cytometry. (f) Quantitative analysis of C11-BODIPY fluorescence intensity. (g) Cell viability detected using a CCK-8 kit. (h) Cell mitochondrial membrane potential detected using JC-1 staining and flow cytometry. (i) Quantitative analysis of (h). (j) Cell death ratio detected using PI staining and flow cytometry. (k) Quantitative analysis of (j). n = 3 per group. Data are presented as the mean ± SEM. ^∗P < 0.05; ^∗∗P < 0.01.