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. 2020 Feb 7;26(6):809–819. doi: 10.1093/ibd/izaa015

FIGURE 6.

FIGURE 6.

DNMT1 mediates the silencing of miR-199a-5p in fibrostenotic Crohn’s disease. A, Expression of miR-199a-5p increased in SEMF after treatment with the DNMT1 inhibitor, 5’-Azacytidine (5’Aza, 5 μg/mL) for 16 hours. B, The ratio of methylated/unmethylated miR-199a-5p decreased in SEMF from fibrostenotic ileum of patients treated with 5’Aza. C, Expression of TGFB1 and COL 1A1 RNA transcripts were measured by qRT-PCR and decreased in SEMF of fibrostenotic ileum after treatment with 5’Aza. D, Representative Western blot and densitometric analysis showing that in normal ileal SEMF from patients with fibrostenosis knockdown of DNMT1 resulted in lower GRP78 and collagen 1A1 production compared with SEMF transfected with scramble siRNA. Results represent the mean ± SEM of 5 to 6 experiments using normal SEMF isolated from 5 to 6 patients with fibrostenotic Crohn’s disease. *Denotes P < 0.05 between vehicle and 5’Aza treated cells or between scramble siRNA and siRNA DNMT1 transfected cells. #Denotes P < 0.05 between scramble siRNA and scramble siRNA + tunicamycin (TM). **Denotes P < 0.05 between scramble siRNA + TM and siRNA DNMT1 + TM.