Figure 5 | Scientific Reports

Figure 5

From: Chronic stress and intestinal barrier dysfunction: Glucocorticoid receptor and transcription repressor HES1 regulate tight junction protein Claudin-1 promoter

Figure 5

Dynamic expression of CLDN1, HES1 and NR3C1 and protein-DNA interaction in Caco-2/BBe cells during 21-day differentiation period. (a) Fresh medium supplied with vehicle or cortisol (500 nM) were added to the plates 24 h after Caco-2/BBe cells were seeded and then replaced every 3 days. Cells were harvested after 2 h cortisol treatment or 2, 6, 11, 21 days after seeding. High (H) exposure and low (L) exposure of X-ray films during Western blot were conducted in order to show the complete dynamic range of protein expression levels. (n = 3; two-tailed test; *P < 0.05; **P < 0.01) (Pre-cropped blots are included in a Supplementary Information). (b) ChIP analysis of human CLDN1 promoter region using HES1 and NR3C1 antibodies. Binding of transcription suppressor HES1 and transcription activator NR3C1 to CLDN1 promoter showed pattern correlated with CLDN1 protein levels. NR3C1 binding to the CLDN1 promoter is non-detectable at day 1 whereas NR3C1 protein level is the lowest. HES1 binding to the CLDN1 promoter is non-detectable at day 11 whereas HES1 level is the lowest (P < 0.01, n = 3).

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