Figure 5.

Detection of mTOR in mitochondria. a, presence of activated mTOR in mitochondria was detected by proximity ligation assay using phosphorylated mTOR and DNA polymerase γ antibodies. Briefly, the cells were treated with UVB radiation, and after 1 h cells were fixed with 4% formalin, then treated with two specific antibodies with different origins, and then probed with Duo-linked (PLA kit) secondary antibody according to the manufacturer's instructions. b, presence of mTORC2 in mitochondria was detected in purified mitochondria. First, mitochondria were isolated using mitochondria purification kit and subjected to Western blotting to identify the mTOR component in purified mitochondrial fraction. c, increase in activated mTOR was detected in UVB-treated mitochondria compared with control. Pretreatment with MnP inhibits mTOR phosphorylation (Ser-2481), AKT phosphorylation (Ser-473), and rictor levels in mitochondria. Statistical significance is indicated by asterisks: *, p ≤ 0.05, and **, p ≤ 0.01.