Fig. 5

IGF2 knockdown in AML12 cells impairs mitochondrial functions. AML12 cells were transfected with sh-IGF2 or its corresponding negative control sh-NC treated with d-gal or H₂O₂. Representative oxygen consumption rates in AML12 cells treated with d-gal (A) or H₂O₂ (C). Basal and maximal respiration, proton leakage, ATP production and spare respiratory capacity in AML12 cells treated with d-gal (B) or H₂O₂ (D). Representative images for ROS production assessed via flow cytometric analyses, and the quantified ROS production are presented in AML12 cells treated with d-gal (E) or H₂O₂ (F). MMP was tested by flow cytometer, and the quantified MMP are presented in AML12 cells treated with d-gal (G) or H₂O₂ (H). I Mitochondrial ROS assessed by MitoSOX staining in AML12 cells treated with d-gal or H₂O₂. J Representative electron micrographs in AML12 cells treated with d-gal or H₂O₂. K The ratio of NAD⁺/NADH in AML12 cells treated with d-gal or H₂O₂. L Western blotting analysis for protein levels of P53, P21 and P16. The quantitative data are presented. Data are shown as means ± SEM. Student t tests was used for comparison between two groups. One-way ANOVA was used for comparison among multiple groups. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. All experiments were repeated three times independently