Fig. 4
Mitochondrial leak state respiration is elevated in Tug1 KD myoblasts and differentiating myotubes. A C2C12 cells were transfected with antisense LNAs (10 nM) for Tug1 knockdown (Tug1 KD) or control then proliferated (Prolif) for 24 h or differentiated (Diff) for 1, 2 or 6 days prior to high-resolution mitochondrial respiratory analyses. B Representative trace showing O2 flux for control (blue) overlaid with Tug1 KD (red) throughout a substrate, uncoupler and inhibitor titration (SUIT) protocol. C Quantification of O2 consumption during specific respiratory states (Leak, non-phosporylating O2 flux; OXPHOS, ADP stimulated oxidative phosphorylation; ETS, uncoupled electron transport system) supported by complex I linked (malate + glutamate) and complex II linked (succinate) substrates, normalised to total cellular protein. Data are mean (SD) for n=3 independent experiments for each time point, analysed by ordinary two-way ANOVA for main effects of LNA and time, with Bonferroni post hoc tests for significant interaction effects: *p<0.05 Tug1 KD compared to NC. See also Additional file 1: Supplementary Fig. S4; individual data are available at DOI: 10.6084/m9.figshare.20175770 [24]