1. Volume 121, Issue 4, Pages 915-930 (October 2001)
Somatostatin suppresses endothelin-1–induced rat hepatic stellate cell contraction via somatostatin receptor subtype 1 
Hendrik Reynaert, Freya Vaeyens, Hong Qin, Karine Hellemans, Nirjhar Chatterjee, Dominique Winand, Erik Quartier, Frans Schuit, Daniel Urbain, Ujendra Kumar, Yogesh C. Patel, Albert Geerts 
Gastroenterology 
Volume 121, Issue 4, Pages (October 2001)
DOI: /gast
Copyright © 2001 American Gastroenterological Association Terms and Conditions

2. Fig. 1
(A) Expression of SSTRs in spleen (lane 1) and rat total liver (lanes 2–5). Lanes 2–5 represent RT-PCR reactions performed on cDNA derived from PolyA+ RNA for 31, 34, 37, and 40 cycles using SSTR subtype–specific primers. Actin primers were used as a control to check for genomic DNA contamination. Primers amplified fragments corresponding to SSTR1, SSTR2, and SSTR3 when PCR reactions were performed on cDNA obtained from total liver. Signals appeared at 37 cycles and became stronger at 40 cycles. Actin primers amplified 1 fragment of 300 nucleotides, showing that only cDNA was amplified. (B) Expression of SSTRs in spleen (lane 1) and cultured rat HSCs (lanes 2–5). Specific primers were found to generate amplicons corresponding to SSTR1, SSTR2, and SSTR3 when 37, 34, and 40 cycles, respectively, were applied. Actin primers amplified 1 fragment of 300 nucleotides. (C) Sequencing results of amplified RT-PCR products from SSTR1-3. Primers are underlined, and bases in bold represent differences from the sequences in the literature. Homology was 99% for SSTR1, 100% for SSTR2, and 98% for SSTR3.
Gastroenterology  , DOI: ( /gast )
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3. Fig. 2
Immunoreaction of anti-SSTR antisera to their corresponding antigenic peptides was measured in triplicate by direct ELISA. Fifty percent effective dilutions were approximately 1/6400 for SSTR 1 and 2 antisera and 1/12,800 for SSTR 3 antiserum. Results are expressed as means ± SD.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Western blot analysis of SSTR1–3 antisera. (A) Diluted SSTR1, SSTR2, and SSTR3 antisera (1/50) were preincubated with their corresponding peptides in increasing doses before Western blotting was performed on protein extracts of cultured rat HSCs. Lane 1, 500 μg/mL; lane 2, 5000 μg/mL; lane 3, 10,000 μg/mL; lane 4, 50 μg/mL; lane 5, 150 μg/mL; lane 6, 450 μg/mL; lane 7, 45 μg/mL; lane 8, 450 μg/mL; lane 9, 4500 μg/mL. (B) Western blot of SSTR1, SSTR2, and SSTR3 on HSCs and pituitary, used as positive control tissue. (C) Deglycosylation of SSTR1–3 with increasing doses of N-glycosidase F. Lane 1, 0 U/10 μg total protein; lane 2, 0.3 U/10 μg total protein; lane 3, 0.6 U/10 μg total protein; lane 4, 1.2 U/10 μg total protein.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Immunohistochemical analysis of SSTR subtypes 1, 2, and 3 in rat liver. Stellate cells with typical fat droplets and long cytoplasmatic processes penetrating between hepatocytes are marked with arrows. (A–D) Normal rat liver is shown in column 1; (E–H) rat liver exposed to CCl4 for 4 weeks is represented in column 2. (A, E) In negative control sections, the primary antibody was replaced by nonimmune serum. No immunoreaction was observed. (B–D) In normal rat liver, stellate cells are not stained by the 3 different SSTR subtype–specific antibodies. (F–H) In CCl4-treated animals, however, very intense immunoreaction occurred for the 3 antibodies. SSTR subtypes 1 and 2 were present in intralobular and septal stellate cells, whereas SSTR subtype 3 was confined to septal cells (original magnification 200×).
Gastroenterology  , DOI: ( /gast )
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6. Fig. 5
Hydrated collagen lattices, photographed 3 hours after addition of vasoactive substances. (A) Buffer only; (B) 10−8 mol/L SST; (C) 2 × 10−8 mol/L ET-1 only; (D) addition of 2 × 10−8 mol/L ET-1 15 minutes after preincubation of the collagen gel with 10−8 mol/L SST; (E) addition of 2 × 10−8 mol/L ET-1 15 minutes after preincubation of the collagen gel with 10−8 mol/L SST.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

7. Fig. 6
(A) Mean percent gel surface area of collagen gels, compared with initial surface area, after incubation with buffer, 10−8 mol/L SST, 2 × 10−8 mol/L ET-1, and 10−8 mol/L SST for 15 minutes followed by 2 × 10−8 mol/L ET-1 for 3 hours and ET-1 for 15 minutes followed by 10−8 mol/L SST for 3 hours (*P < 0.05). (B) Mean contraction of collagen gels exposed to the combination of 10−8 mol/L SST and 2 × 10−8 mol/L ET-1 after 3 hours compared with the contraction of cells exposed to 2 × 10−8 mol/L ET-1 alone (*P < 0.05).
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

8. Fig. 7
Collagen gels photographed 3 hours after addition of vasoactive substances. (A) Buffer only; (B) 2 × 10−8 mol/L ET-1 only; (C–E) combination of 2 × 10−8 mol/L ET-1 and 10−8 mol/L of L (SSTR subtype 1 agonist), L (SSTR subtype 2 agonist), and L (SSTR subtype 3 agonist), respectively.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

9. Fig. 8
Average contraction of gels exposed for 3 hours to the combination of 10−8 mol/L L , L , and L and 2 × 10−8 mol/L ET-1 relative to the contraction of cells exposed to buffer and ET-1 alone (n = 3). Bars represent mean ± SD percent contraction (*P < 0.05).
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions