Fig. 5

S-Equol regulates Chrebp/Txnip signaling in high glucose cultured INS-1 cells. INS-1 cells were treated for 48 h with or without S-Equol (0.1, 1 and 10 μM) in the continuous presence of high glucose (26.2 mM), and cells cultured with 5 mM of glucose denote the control. (a) Western blotting analysis to determine Txnip, Chrebp and Mlx, and (b) densitometric quantification of the relative folds of the proteins /actin ratio in INS-1 cells was shown in bar graph; (c) Quantitative RT-PCR analysis of Txnip-mRNA transcription; (d) Txnip promoter activities. Luciferase activity was expressed as the percentage relative to the control group; (e) ChIP assay. Cell lysate was precipitated with anti-Chrebp and normal IgG was used as a negative control. (f - h) The effect of si-Chrebp on protein levels of Txnip and Chrebp. Each value represents the amount of protein relative to that of the control group transfected with the scrambled siRNA in the same set of experiments. Three to five independent experiments were run in each experiments and results were presented as means ± SD. *P < 0.05 vs. the Control group, ^# P < 0.05 vs. the High glucose group