Figure 7.

Exosomal circHIPK3 derived from hypoxia-pretreated CMs induces protection from oxidative injury via miR-29a/IGF-1 in CMVECs. (a) Representative dot plots of cell apoptosis after Annexin V/PI dual staining are shown. (b) The percentage of apoptotic cells represents both early and late apoptotic cells; n = 3. (c) The intracellular ROS level was determined by FCM. The P2 percentage indicates the proportion of cells with increased ROS production, with signals above background 2′,7′-dichlorofluorescein (DCF) fluorescence levels. (d) Quantitative analysis of ROS levels; n = 3. (e) Representative immunofluorescence staining of TUNEL (green) and DAPI (blue) staining and merged images. Photos were randomly captured using a fluorescence microscope. Scale bar = 20 μm. (f) The panel shows the percentage of TUNEL-positive cells; n = 6. (g) Graph represents SOD levels; n = 9. (h) Graph represents MDA levels; n = 9. (i) Apoptosis-related genes, such as procaspase-3, cleaved caspase-3, Bax, and Bcl-2, were detected by immunoblotting. (j, k) Quantitative analysis of the apoptosis-related proteins; n = 3. (l) qRT-PCR analysis of miR-29a expression in CMVECs after different treatments. (m) qRT-PCR analysis of IGF-1 mRNA expression in CMVECs after different treatments. (n) IGF-1 protein was detected by immunoblotting. (o) Quantitative analysis of IGF-1 protein; n = 3. ^∗P < 0.05 compared with the H₂O₂ group; ^#P < 0.05 compared with the sicircHIPK3-exos group.