1. Volume 119, Issue 1, Pages 211-219 (July 2000)
Cytokine regulation of chemokine (IL-8, MCP-1, and RANTES) gene expression in human pancreatic periacinar myofibroblasts 
Akira Andoh, Hiroki Takaya, Takao Saotome, Mitsue Shimada, Kazunori Hata, Yoshio Araki, Fumiyasu Nakamura, Yutaka Shintani, Yoshihide Fujiyama, Tadao Bamba 
Gastroenterology 
Volume 119, Issue 1, Pages (July 2000)
DOI: /gast
Copyright © 2000 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Effects of various cytokines on IL-8 and MCP-1 secretion by human pancreatic periacinar myofibroblasts. The cells were incubated with various cytokines (50 ng/mL, except IFN-γ [200 U/mL]) for 48 hours, and IL-8 and MCP-1 levels were determined by ELISA. Values are expressed as means of quadruplicate cultures.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

3. Fig. 2
IL-1β– and TNF-α–induced chemokine secretion in human pancreatic periacinar myofibroblasts. The cells were incubated with increasing concentrations of IL-1β and TNF-α for 48 hours, and (A) IL-8, (B) MCP-1, and (C) RANTES levels were determined by ELISA. Values are expressed as means ± SD of quadruplicate cultures.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Northern blot analysis of chemokine mRNA expression in human periacinar myofibroblasts. The cells were cultured in the absence or presence of IL-1β (50 ng/mL) or TNF-α (50 ng/mL) for 4 hours, and total cellular RNA was extracted. Prolonged exposure of the membrane to x-ray film was needed to detect RANTES mRNA expression. Ribosomal RNA, stained by ethidium bromide, is shown in the lower part.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Kinetics of chemokine mRNA expression in human pancreatic periacinar myofibroblasts. (A) The cells were incubated with IL-1β (50 ng/mL) and (B) TNF-α (50 ng/mL), and the accumulation of IL-8, MCP-1, and RANTES mRNA was sequentially determined by Northern blotting. The radioactivity of each band was quantified by the Instant Image Electronic Autoradiography System (Packard), and relative radioactivity was compared with the radioactivity of unstimulated cells.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Nuclear run-on assays for evaluation of transcriptional activation of chemokine genes. Nuclei were isolated from periacinar myofibroblasts 3 or 12 hours after stimulation with IL-1β or TNF-α, and nuclear run-on assays were performed in the presence of [α-32P]uridine triphosphate, as described in Materials and Methods.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Evaluation of chemokine mRNA stability. Cells were stimulated with TNF-α (50 ng/mL) for 12 hours and treated with actinomycin D (5 μg/mL) for various time periods. Chemokine mRNA expression was detected by Northern blot and analyzed by the Instant Image Electronic Autoradiography System (Packard).
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

8. Fig. 7
Electrophoretic gel mobility shift assays for NF-κB and NF-IL6 DNA-binding activities. Periacinar myofibroblasts were incubated with medium alone, IL-1β (50 ng/mL), or TNF-α (50 ng/mL) for 1.5 hours, and the nuclear extracts were prepared. (A) NF-κB. Lane 1, medium alone; lane 2, IL-1β (50 ng/mL); lane 3, IL-1β + anti-p50 antibody; lane 4, IL-1β + anti-p65 antibody; lane 5, IL-1β + cold probe; lane 6, TNF-α (50 ng/mL); lane 7, TNF-α + anti-p50 antibody; lane 8, TNF-α + anti-p65 antibody; lane 9, TNF-α + cold probe. Dashed arrows indicate nonspecific DNA-protein bindings. (B) NF-IL6. Lane 1, medium alone; lane 2, IL-1β (50 ng/mL); lane 3, IL-1β + cold probe; lane 4, IL-1β + anti-NF-IL6 antibody; lane 5, TNF-α (50 ng/mL); lane 6, TNF-α + cold probe; lane 7, TNF-α + anti–NF-IL6 antibody.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions

9. Fig. 8
Effects of the NF-κB inhibitor PDTC and TPCK on IL-1β– or TNF-α–induced chemokine mRNA expression. The cells were incubated with IL-1β (50 ng/mL), IL-1β (50 ng/mL) + either PDTC (20 μmol/L) or TPCK (20 μmol/L), TNF-α (100 ng/mL), or TNF-α + either PDTC (20 μmol/L) or TPCK (20 μmol/L). Next, total cellular RNA was extracted. IL-8 and MCP-1 mRNA expression was analyzed by Northern blot after 3-hour stimulation, and RANTES mRNA expression was analyzed after 6-hour stimulation.
Gastroenterology  , DOI: ( /gast )
Copyright © 2000 American Gastroenterological Association Terms and Conditions