Journal: Scientific Reports

Article Title: Molecular hydrogen suppresses activated Wnt/β-catenin signaling

doi: 10.1038/srep31986

Figure Lengend Snippet: (a) HCS-2/6 human chondrosarcoma cells were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 24 h. The Wnt/β-catenin signaling activity was measured by Topflash luciferase reporter assay ( n = 9). * P < 0.05 and ** P < 0.01 by Student’s t -test. (b) Human OAC cells were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 24 h. Representative Western blots are shown with densitometry of β-catenin/β-actin ( n = 4). * P < 0.05 by Student’s t -test. (c–g) Human OAC cells (clone 1) were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 24 h. Expression of AXIN2 ( n = 3) (c) , SOX9 ( n = 6) (d) , MMP3 ( n = 6) (e) , COL2A1 ( n = 3) (f) , and ACAN ( n = 3) (g) were quantified by qRT-PCR. * P < 0.05 by Student’s t -test. (h) Differentiated ATDC5 cells were treated with control CM (Cont.), Wnt3a CM, or 2 μM BIO with 10% H 2 or 10% N 2 gas for 48 h. Proteoglycans were stained with Alcian blue (upper panel) and quantified by measuring the optical intensity at 630 nm of the cell lysates ( n = 9 for Cont.-treated and BIO-treated cells, n = 6 for Wnt3a-treated cells). * P < 0.05 by Student’s t-test. Non-significant P values less than 0.10 are indicated above each pair.

Article Snippet: HepG2 and ATDC5 cells were from RIKEN BioResource Center.

Techniques: Control, Activity Assay, Luciferase, Reporter Assay, Western Blot, Expressing, Quantitative RT-PCR, Staining