fig6

Figure 6. LINC01133 promotes cisplatin resistance by maintaining mitochondrial structural and functional integrity through calcium homeostasis. (A) Effect of LINC01133 expression on the intracellular calcium ion concentration detected using the fluorescent probe Fluo-4 AM; (B) Effect of LINC01133 expression on the mitochondrial calcium ion concentration detected using the Rhod-2 AM fluorescent probe; (C) Role of LINC01133 expression in determining the mitochondrial calcium ion distribution using fluorescence colocalization imaging. Hoechst 33342 was added for nuclear staining (blue), MitoTracker was used for mitochondrial labeling (green), and Rhod-2 AM was used for mitochondrial calcium labeling (red). Scale bar: 50 μm; (D) Role of LINC01133 expression in mitochondrial ultrastructure determined through TEM. Scale bar: 5 μm; (E) Effect of LINC01133 expression on COX IV and TOMM20 protein levels detected by Western blotting. Densitometric values of the target proteins were normalized to GAPDH and are presented as relative protein expression levels. Data were obtained from three independent biological replicates and are presented as the mean ± SD (n = 3); (F) Effect of LINC01133 expression on mitochondrial membrane potential assessed using JC-1 staining; (G) Effect of LINC01133 expression on mitochondrial ROS levels detected using MitoSOX staining; (H) Effect of LINC01133 expression on ATP content measured using a chemiluminescence assay; (I) Effect of LINC01133 expression on mtDNA copy number determined by PCR. One-way ANOVA was employed to compare several groups. ^***P < 0.001. TEM: Transmission electron microscopy; SD: standard deviation; ROS: reactive oxygen species; ATP: adenosine triphospate; mtDNA: mitochondrial DNA; PCR: polymerase chain reaction; ANOVA: analysis of variance; DDP: cisplatin.