Fig. 3.

Cellular respiration and oxidative metabolism: (A–D) Cellular oxygen consumption rates (OCR), determined by Seahorse XFp Extracellular Flux analyser. Mitochondrial inhibitors/uncouplers were added sequentially during the experiment in the following concentrations: oligomycin 1 μM (O), FCCP 1 μM (F) and antimycin A/rotenone 0.5 μM (AA/R). MSTN genotypes; CC/II (green), CT/IN (blue) and TT/NN (red); (A) OCR of primary equine skeletal muscle cells; (B) maximal respiration (maximum rate measurement after FCCP injection) – (non-mitochondrial respiration after AA/R injection) primary equine skeletal muscle cells; (C) OCR of immortalised equine skeletal muscle cells; - and + CoQ₁₀ (5 μM for 24 h); (D) maximal respiration immortalised equine skeletal muscle cells, - (plain bars) and + CoQ₁₀ (5 μM for 24 h) (dotted bars). (E–G) FLIM analysis of the immortalised cell lines (NAD(P)H-FLIM imaging; marker of oxidative metabolism); (E) average lifetime (ns); (F) optical redox ratio (a.u.); (G) NAD(P)H-FLIM images, each image is representative of 3 independent experiments. Results presented with mean ± SEM (N = 3 per genotype, performed in triplicate technical replicates), p-values where shown indicate significance as measured by; a one-way (B, E, F) or two-way (D) ANOVA with a Tukeys multiple comparisons test * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)