Journal: Nature Communications

Article Title: Transcriptomic profiling of skeletal muscle adaptations to exercise and inactivity

doi: 10.1038/s41467-019-13869-w

Figure Lengend Snippet: a Genes significantly modified by acute aerobic and resistance exercise and inactivity were overlapped in a Venn Diagram. b DNAJA4 , KLHL40 , NR4A3 , and VGLL2 were validated in an independent cohort of pre- and post-acute aerobic exercise. Individual paired t -tests vs pre, n = 8 biologically independent volunteers, ** p < 0.01. c DNAJA4 , NR4A3 , KLHL40 , and VGLL2 gene expression following electrical pulse stimulation in primary human myotubes. Individual paired t -tests vs basal, n = 8 biologically independent primary cells from different donors, ** p < 0.01.

Article Snippet: Human primary myotubes transfected by either a siRNA against NR4A3 or a scrambled siRNA were subjected to electrical pulse stimulation (C-Pace EP, Ionoptix) at 40 V, 2 ms, 1 Hz during 3 h in low glucose serum-free DMEM.

Techniques: Modification, Gene Expression

Journal: Nature Communications

Article Title: Transcriptomic profiling of skeletal muscle adaptations to exercise and inactivity

doi: 10.1038/s41467-019-13869-w

Figure Lengend Snippet: a N R4A3 responds in a time- and intensity-dependent manner to electrical pulse stimulation. Data are mean ± SEM, n = 3 biologically independent primary cells from different donors, two-way ANOVA (time, intensity), *overall effect p < 0.05. b Silencing efficiency using siRNA against NR4A3, n = 4 biologically independent primary cells from different donors, individual paired t -test vs scramble, *** p < 0.001. c Electrical pulse stimulation-induced glucose uptake is abolished after NR4A3 silencing. Two-way ANOVA (si NR4A3 , EPS), n = 6 biologically indepe n dent primary cells from different donors, * p < 0.05. d Silencing NR4A3 using siRNA modifies the mRNA levels of exercise- and inactivity-responsive genes. Data are mean ± SEM, n = 4 biologically independent primary cells from different donors, individual paired t -test vs scramble. e Silencing of NR4A3 correlates with inactivity observed in MetaMEx. f Reduction of NR4A3 level impairs basal and maximal oxygen consumption measured by Seahorse XF analysis. Individual paired t -tests vs scramble, n = 5 biologically independent primary cells from different donors, * p < 0.05, ** p < 0.01. g Silencing of NR4A3 leads to a drift of muscle cells towards a more quiescent phenotype. Data are mean ± SEM. h Silencing of NR4A3 decreases the abundance of mitochondrial complexes. Representative blot and quantification, n = 6 biologically independent primary cells from different donors, two-way ANOVA, significant effect of silencing ( p = 0.018), * p < 0.05 uncorrected Fisher’s LSD post-test. i The increase in glycolysis (ECAR) induced by beta-adrenergic stimulation (20 µM salbutamol for 3 h) is impaired in the absence of NR4A3 . SeaHorse XF experiment, two-way ANOVA (si NR4A3 , Salbutamol), n = 7 biologically independent primary cells from different donors, ** p < 0.01. AA: acute aerobic, AR: acute resistance, IN: inactivity, TA: training aerobic, TR: training resistance.

Article Snippet: Human primary myotubes transfected by either a siRNA against NR4A3 or a scrambled siRNA were subjected to electrical pulse stimulation (C-Pace EP, Ionoptix) at 40 V, 2 ms, 1 Hz during 3 h in low glucose serum-free DMEM.

Techniques: