Fig. 5: The expression of DDAH1 is responsive to mechanical force via TAZ/SMAD4 signaling pathway. | Nature Communications

Fig. 5: The expression of DDAH1 is responsive to mechanical force via TAZ/SMAD4 signaling pathway.

From: Mechanical force promotes dimethylarginine dimethylaminohydrolase 1-mediated hydrolysis of the metabolite asymmetric dimethylarginine to enhance bone formation

Fig. 5

a The typical images of immunofluorescence staining of DDAH1 (green) and DDAH2 (red) in bones of exercise-treated or tail-suspension mice. Scale bar = 100 μm. b Quantification analysis of DDAH1 and DDAH2 expression of bone parameters. ns no significance. Ground group mice, n = 5. Unload group mice, n = 5. Control group, n = 5. Exercise group, n = 5. *p < 0.05. ***p < 0.005. Data are represented as mean values ± SD. c Metabonomics of serum samples of control mice and exercise mice. d Volcano plot of different metabolites responsive to exercise treatment. e Serum samples of exercise mice or tail-suspension mice were analyzed by LC-MS assay. Ground group mice, n = 5. Unload group mice, n = 5. Control group, n = 5. Exercise group, n = 5. **p < 0.01. ***p < 0.005. Data are represented as mean values ± SD. f Western blotting analysis of the levels of phosphor-YAP, YAP, TAZ, ALP, and DDAH1 in osteoblasts treated with tension force for 6 h or not, and then cultured for 18 h, the cycle was repeated three times. g Co-IP assay of the association between SMAD4 and TAZ in simulation with tension force. h Representative immunofluorescence staining images of the association between SMAD4 (green) and TAZ (red) in simulation with tension force. Scale bars = 100 μm. i ChIP assay of SMAD4 clustering on the promoter regions of Ddah1 in stimulation with tension force. j Quantification analysis of ChIP assay. Control, n = 3. Tension, n = 3. *p < 0.05. **p < 0.01. Data are represented as mean values ± SD. k Luciferase reporter genes assay of Ddah1 from WT primary osteoblasts in stimulation with tension force by the silence of TAZ and SMAD4. All groups, n = 3. *p < 0.05. **p < 0.01. ***p < 0.005. Data are represented as mean values ± SD. The data shown in all panels were analyzed by an unpaired two-tailed Student’s t test. One-way analysis of variance (ANOVA) with post-hoc Tukey’s test was used for experiments with three or more groups.

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