Fig. 2
Mutant mitochondrial tRNATrp triggers widespread mitochondrial dysfunction. a Enzymatic activities of individual mitochondrial respiratory chain complexes for cultured myoblasts of both controls (n = 10, closed circles) and the patient (open circles). Error bars indicate as the means with SD of controls. All samples were measured in triplicate and averaged. b Representative images of cytochemical staining of COX and SDH for cultured myoblasts from both controls and the patient. Cell nuclei were co-stained with hematoxylin. c,d Immunodetection of c individual respiratory chain complexes and d respiratory chain supercomplexes for isolated mitochondria from cultured myoblasts of both controls and the patient. e Gene expression of several CIV structural subunits for extracted mRNA from cultured myoblasts of both controls and the patient. f Protein expression of several CIV structural subunits for isolated mitochondria from cultured myoblasts of both controls and the patient. g Representative images of immunocytochemical staining of CIV structural subunits for cultured myoblasts of both controls and the patient; MT-CO1 (red), COX4 (red). Mitochondria were co-stained with MitoTracker (green). h ATP level for cultured myoblasts of both controls (n = 10, closed circles) and the patient (open circles). Error bar indicates as the mean with SD of controls. All samples were measured in triplicate and averaged. i Oxidative stress level for cultured myoblasts of both controls (n = 10, closed circles) and the patient (open circle). Fluorescence intensity ratio of MitoSOX/MitoTracker served as mitochondrial ROS level. Error bar indicates as the mean with SD of controls. All samples were measured in triplicate and averaged. j Membrane potential (ΔΨm) level for cultured myoblasts of both controls (n = 10, closed circles) and the patient (open circle). Fluorescence intensity ratio of JC-1 dye aggregates/monomers served as mitochondrial ΔΨm level. Error bar indicates as the mean with SD of controls. All samples were measured in triplicate and averaged