Fig. 5.

Effects of pro-oxidants on AMPK activity and Cn pathay. AMPKα (Thr172) activity in Raw264.7 and C2C12 cells were measured following treatment with DTPP ⁺ control or TPP + conjugated agents (MitoPQ and MitoMet) for 16 h at the indicated doses. A, Phosphorylated AMPKα level was measured in Raw264.7 cells and C2C12 cells. B, immunoblot analysis of phosphor-AMPKα protein along with GAPDH as loading control. The Bar diagram representing (the band intensity AMPKα Phos/tolal) The blots are representative of three different runs (n = 3). C-E, mRNA expression profile of different MtRS signaling markers at different time of hypoxia in HCT116 cells. The mRNA levels were monitored in qRT-PCR assays. The results from 3 independent experiments and normalized to beta actin mRNA levels and expressed as a fold change over the normoxic control. F, Matrigel invasion of control HCT116 cells treated with or without mito-targeted agents for 16h, before cells were loaded in to the chamber. G, quantitation of invading cells from triplicate experiments as in F. Significance was calculated by one-way ANOVA with Tukey's multiple-comparison test, and data are presented as treatment group versus no-treatment control. In all experiments, error bars represent standard deviations (*p < 0.05, **p < 0.01).