Fig. 3: Recombinant CLCF1 treatment stimulates glucose uptake and glucose metabolism.
From: Exercise-induced CLCF1 attenuates age-related muscle and bone decline in mice
A, B Oxygen consumption rate (OCR) and quantification of basal respiration, ATP production, and maximal respiration in C2C12 myotubes treated with CLCF1 (100 ng/ml) for 48 h (vehicle, n = 10; CLCF1, n = 9; biological replicates, p = 1.0e-6, 7.0e-7, 1.3e-8). C OCR traces and D quantification in response to UK5099. (n = 10; p = 1.6e-5, 5.2e-7, 1.6e-4, 4.6e-5, 2.0e-11, 2.0e-11, 2.0e-11). E, F Mitochondrial DNA content and expression of mitochondrial genes in C2C12 myotubes after 48 h of CLCF1 treatment (n = 3). G Relative level of glucose uptake in C2C12 myotubes with CLCF1 treatment overnight (n = 6; p = 9.8e-3, 2.86e-2). H Relative level of fatty acid uptake in C2C12 myotube with CLCF1 overnight (n = 5). I Left: Western blot analysis of GLUT1 and GLUT4 in membrane and cytosolic fractions after 24 h of treatment with vehicle or CLCF1 in C2C12 myotubes (n = 3; p = 0.0469). Right: Quantification of Western blots was performed by densitometry analysis. J Measurement of glycolysis by extracellular acidification rate (ECAR) in C2C12 myotubes treated with vehicle or CLCF1 for 48 h (n = 8). K Quantification of glycolysis and glycolytic capacity (p = 3.0e-4, 2.0e-4). L Expression of glycolysis-related enzymes in C2C12 myotubes treated with vehicle or CLCF1 for 24 h. M Quantification of glycolytic enzymes (n = 4; p = 3.44e-2, 4.6e-3, 2.05e-2, 2.2e-3). N Top: Expression of glycolysis-related enzymes in muscle of young (3 mo, male), old (20 mo, male), and old-CLCF1 (20 mo, male). Bottom: Quantification of western blots (n = 4; p = 0.0313, 0.0135, 0.0049, 0.0001). O Intraperitoneal glucose tolerance test (IPGTT) in young, old, and old-CLCF1 (p = 4.8e-5, 7.5e-5). P Area under curve (AUC) (young, n = 12; old, n = 10; old_CLCF1, n = 8; biological replicates, p = 0.0149, 0.0293). Q Insulin tolerance test in young, old, and old-CLCF1 (p = 2.5e-6, 8.2e-7). R AUC young, n = 8; old, n = 11; old_CLCF1, n = 10; p = 0.0251, 0.0135. Data are presented as means ± s.e.m.; two-tailed unpaired Student’s t-test (B, E–I, K, M); one-way ANOVA with post hoc Bonferroni’s multiple comparison test (J, N, P, R); or two-way ANOVA with post hoc Tukey’s multiple comparison test (D, O, Q). *p < 0.05, **p < 0.005 and ***p < 0.001; n.s., p > 0.05. Source data are provided as a Source Data file.
