Journal: The Journal of Physiology

Article Title: Forkhead BoxO transcription factors restrain exercise-induced angiogenesis

doi: 10.1113/jphysiol.2014.275867

Figure Lengend Snippet: A, FVB/n mice (n = 4) exercised for 1 day or remained sedentary (S). Muscles were extracted immediately (Ex), or 2 h after training (Ex+2 h). B, FVB/n FoxOL/L mice (n = 6) trained for 1, 7, 10 or 14 days (D1, D7, D10, D14) or remained sedentary (S), and all muscles were extracted 2 h after the final training session. In both, THBS1 protein was assessed relative to β-actin. For densitometric analysis of THBS1, intensities of the three bands evident at ∼ 175 kDa were summed together. *P < 0.05 vs. sedentary; ##P < 0.01 vs. Ex (Tukey post hoc analysis).

Article Snippet: Membranes were probed using antibodies as follows: FoxO1, FoxO3a, p-Thr24 FoxO1/p-Thr32 FoxO3a, total AKT1/2/3, p-Ser473 Akt, total JNK1, pThr183/Tyr185 JNK, α/β-Tubulin (1:1000; #9454, 2497, 9464, 9272, 4058, 9258, 9255, 2148, respectively; Cell Signalling, Pickering, ON, Canada), THBS1 (1:200; #G3685605 Cedarlane Laboratories, Burlington, ON, Canada), Histone 2B (1:1000; #ab64165 Abcam, Cambridge, MA, USA) and β-actin (1:5000; #47778 Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Muscles

Journal: The Journal of Physiology

Article Title: Forkhead BoxO transcription factors restrain exercise-induced angiogenesis

doi: 10.1113/jphysiol.2014.275867

Figure Lengend Snippet: A, FoxO1 and FoxO3a protein levels were analysed in endothelial cells isolated from epididymal fat of FoxOL/L and FoxOΔ mice (n = 3). Representative blots are shown, with the respective densitometric values (relative to β-actin) listed under each blot. B, muscles were extracted from FoxOL/L and FoxOΔ sedentary (S) mice, or 2 h after an acute exercise bout (D1). FoxO1 and FoxO3a protein levels were assessed by Western blot to determine the extent of deletion from whole gastrocnemius extracts, and quantified relative to tubulin. Significant exercise and genotype effects were observed (two-way ANOVA) for both FoxO1 (P = 0.0008 and P = 0.006, respectively) and FoxO3a (P = 0.01 and P = 0.009, respectively). C, FoxOL/L and FoxOΔ mice (n = 6) trained for 1, 7 or 14 days, or remained sedentary (S) and muscles were extracted 2 h after training. THBS1 mRNA levels were measured by qPCR, normalizing to the housekeeping gene HPRT. Significant training (P < 0.0001) and genotype (P < 0.01) effects were observed (two-way ANOVA). D, THBS1 protein was assessed by Western blot of whole gastrocnemius extracts and quantified relative to tubulin. Significant exercise (P < 0.05) and genotype (P < 0.05) effects were observed (two-way ANOVA). *P < 0.05; ***P < 0.001 vs. sedentary FoxOL/L; #P < 0.05; ##P < 0.01; ###P < 0.001 vs. FoxOΔ day 1 (Bonferroni post hoc analysis).

Article Snippet: Membranes were probed using antibodies as follows: FoxO1, FoxO3a, p-Thr24 FoxO1/p-Thr32 FoxO3a, total AKT1/2/3, p-Ser473 Akt, total JNK1, pThr183/Tyr185 JNK, α/β-Tubulin (1:1000; #9454, 2497, 9464, 9272, 4058, 9258, 9255, 2148, respectively; Cell Signalling, Pickering, ON, Canada), THBS1 (1:200; #G3685605 Cedarlane Laboratories, Burlington, ON, Canada), Histone 2B (1:1000; #ab64165 Abcam, Cambridge, MA, USA) and β-actin (1:5000; #47778 Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Isolation, Muscles, Western Blot