Figure 1 | Scientific Reports

Figure 1

From: Molecular hydrogen suppresses activated Wnt/β-catenin signaling

Figure 1

H2 suppresses Wnt/β-catenin signaling in L cells.

(a,b) Cells were treated with control CM (Cont.), Wnt3a CM, 30 mM LiCl, or 2 μM BIO with 10% H2 or 10% N2 gas for 24 h. The Wnt/β-catenin signaling activity was measured by Topflash luciferase reporter assay (n = 4) (a) and expression of Axin2 was quantified by qRT-PCR (n = 3–4) (b). *P < 0.05 and **P < 0.01 by Student’s t-test. (c–e) Cells were treated with pairs of CM (Cont.) and Wnt3a CM (c); 30 mM NaCl and 30 mM LiCl (d); and 0.02% DMSO and 2 μM BIO/0.02% DMSO (e) with 10% H2 or 10% N2 gas for 24 h. Representative Western blots are shown. (f) Cells were treated with 2 μM BIO with 10% H2 or 10% N2 gas for indicated periods of time. The upper panel shows representative Western blots and the lower panel shows densitometry of β-catenin/β-actin (n = 4). #P < 0.05 by two-way repeated measures ANOVA. **P < 0.01 by Student’s t-test with Bonferroni correction for each pair of H2 and N2. (g) Cells were treated with control CM (Cont.), Wnt3a CM, 30 mM LiCl, or 2 μM BIO with either 10% N2 gas, intermittent 10% H2 gas, or continuous 10% H2 gas for 24 h. Representative Western blots are shown with densitometry of β-catenin/β-actin (n = 3). *P < 0.05 and **P < 0.01 by Student’s t-test with Bonferroni correction. (h) Cells were pretreated with 40 μM SP600125 for 30 min. Cells were then added with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H2 or 10% N2 gas for 1 h. Representative Western blots are shown with densitometry of β-catenin/β-actin (n = 4). *P < 0.05 and **P < 0.01 by Student’s t-test.

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