Fig. 3.

Cirsiliol decreases the mitochondrial membrane potential, promotes the generation of ROS, and disrupts mitochondria network morphology. HCT116 and SW480 cells were treated with cirsiliol for 24 h. A, B Mitochondrial membrane potential of HCT116 (A) and SW480 cells (B) was determined by JC-1 staining. Representative fluorescence images of JC-1 aggregate (red) and JC-1 monomer (green) are shown in the Figure. Scale bar: 50 μm. C, D Quantification of mitochondrial membrane potential (JC-1 aggregate/monomer ratio) for HCT116 (C) and SW480 cells (D). E H2DCFDA staining was performed to detect ROS production for HCT116 and SW480 cells. Scale bar: 50 μm. F Quantification of H2DCFDA fluorescence intensity. G We performed Mito-Tracker Red staining to detect the mitochondria network morphology of SW480 cells. Representative images were presented in the figure. Scale bar: 10 μm. H–J MiNA was used to analyze the morphology of the mitochondrial network, including percentage of network mitochondria (H), mean branch length (I), and mitochondrial footprint (J). n = 5 (A-F), and 10 (G–J) per group. Data represent means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 vs cirsiliol 0 μM