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. 2016 Dec 6;187(3):428–440. doi: 10.1111/cei.12896

Figure 4.

Figure 4

Tumour necrosis factor (TNF) induces hypoxia inducible factor‐1α (HIF‐1α) expression through activating nuclear factor (NF)‐кB. (a,b) HT‐29 cells (5 × 105/ml) were stimulated in vitro with interferon (IFN)‐γ (10 ng/ml), interleukin (IL)−17A (10 ng/ml), IL‐23 (10 ng/ml), TNF (10 ng/ml), IL‐10 (10 ng/ml), transforming growth factor (TGF)‐β (10 ng/ml), lipopolysaccharide (LPS) (100 ng/ml), peptidoglycan (PGN) (1 μg/ml), flagellin (100 ng/ml) and polyI:C (10 μg/ml) for 24 h, relative expression of HIF‐1α mRNA was determined by quantitative real‐time–polymerase chain reaction (qRT–PCR) analysis and normalized to glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH). * P < 0·05 versus controls. (c) Relative expression of HIF‐1α mRNA in HT‐29 cells under stimulation with different concentrations of TNF (0, 1, 5, 10, 20, 50 ng/ml) for 24 h. Representative results from three independent experiments are shown. * P < 0·05; ** P < 0·01 versus data cultured in medium alone. (d) Relative expression of HIF‐1α mRNA in HT‐29 cells under stimulation with TNF (10 ng/ml) for 0, 12, 24 and 48 h. Representative results from three independent experiments are shown. ** P < 0·05. (e) Relative expression of HIF‐1α mRNA in colonic tissues from the same Crohn's disease (CD) patients (n = 10) before and after anti‐TNF monoclonal antibody (mAb) treatment [infliximab (IFX), 5 mg/kg]. GAPDH was used as a housekeeping gene. * P < 0·05. (f,g) Freshly isolated colonic biopsies from patients with CD (n = 10) (f) or ulcerative colitis (UC) (n = 10) (g) were cultured in the presence of IFX (50 μg/ml) or control human immunoglobulin (HIg, 50 μg/ml) for 24 h. The levels of HIF‐1α mRNA were determined by qRT–PCR and normalized to GAPDH. * P < 0·05. Data are compiled from three independent experiments. (h−j) HT‐29 cells were treated with TNF (10 ng/ml), TNF (10 ng/ml) and Bay 11‐7082 (1 μM) (h), TNF and SP600125 (10 μM) (i), or TNF and CH‐11 (200 ng/ml) (j) for 24 h, and the levels of HIF‐1α mRNA were determined by qRT–PCR. Representative results from three independent experiments are shown. * P < 0·05.