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. 2013 Mar 20;18(5):631–641. doi: 10.1007/s12192-013-0416-0

Fig. 5.

Fig. 5

Regulation of CBS activity by GC during stress. To determine whether glucocorticoids exerted direct control over cbs transcription, we examined hepatocytes that had been incubated with10−6 mol/l CORT for 1, 6 and 12 h. The expression of GR was increased in nuclei of hepatic cells with CORT treatment (a, top), and was decreased to about 31 % in the hepatic cytoplasm after treatment with CORT for 12 h (a, bottom). Compared with control, GR nuclear translocation was remarkably increased after CORT stress (b). c The level of Sp3 expression in nuclei of GCs-treated hepatocytes was elevated, and Sp1 expression was slightly reduced. As a result, the ratio of Sp1/Sp3 was decreased. Graphical representations of Sp1 and Sp3 are shown in the bottom panel with changes in expression over time. d CBS mRNA levels were markedly lowered in GCs-treated hepatocytes compared with control, to 19 % of the control after CORT treatment for 12 h. e The results from CHIP and PCR showed the numbers of the Sp3 specific fragments (458 bp) were increased after CORT treatment, but the change of Sp1(254 bp) was not observed (data not shown). It suggested that GCs enhance the binding of GR and the regulatory region (GREs) of Sp3 promoter and have no effect on Sp1. β-Actin was used as an internal control, and the change ratio of proteins is normalized by β-actin based on OD value of proteins bands (n = 3, **p < 0.01 compare with control)