1. Distribution of the novel eNOS–interacting protein NOSIP in the liver, pancreas, and gastrointestinal tract of the rat 
Wolfgang Kummer, Peter König 
Gastroenterology 
Volume 123, Issue 1, Pages (July 2002)
DOI: /gast
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2. Fig. 1
Western blot of liver (1), forestomach (2), stomach (3), small intestine (4), large intestine (5), and pancreas (6), 50 μg of protein was loaded for each tissue. Calculated molecular masses of monomeric, dimeric, and tetrameric forms of NOSIP are identified on the right margin.
Gastroenterology  , DOI: ( /gast )
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3. Fig. 2
(A) Esophagus. NOSIP immunoreactivity (IR) was located in the epithelium (ep), the smooth muscle cells of the lamina muscularis mucosae (mm), and the striated muscle cells of the tunica muscularis (m). (B–G) Stomach. NOSIP-IR was found in the epithelium of the forestomach (fs), in gastric glandular cells (gc), and in the muscularis mucosae. Boxed area, see D. (C) In the epithelium of the forestomach, NOSIP-IR was predominantly located in the nuclei of the basal cell layer and in the stratum spinosum. (D) Higher magnification of a region corresponding in location to the boxed area in B. In the glandular cells of the stomach, immediately adjacent to the forestomach, a membrane-bound IR was found. (E) A subset of parietal cells shows nuclear NOSIP-IR (arrows). (F) In the tunica muscularis, the innermost circular muscle cells (cm) and the longitudinal muscle layer (lm) showed intense NOSIP-IR. Between the muscle cells, elongated cells showed NOSIP-IR (arrows). (G, G') Immunolabeling is absent after preabsorption of the antiserum with NOSIP-GST-fusion protein (20 μg/mL). (G') Same area labeled with NOSIP antiserum in an adjacent section depicted in G. (Bars: A, B, F–G' = 100 μm; C–E = 20 μm).
Gastroenterology  , DOI: ( /gast )
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4. Fig. 3
(A, B) Small intestine. (A) NOSIP immunoreactivity was found in the nuclei of enterocytes (arrows) and in smooth muscle cells (small arrows) of the villus stroma. (B) Intense NOSIP immunoreactivity was found in the muscularis mucosae (mm). In the tunica muscularis, NOSIP immunoreactivity was predominantly localized in the innermost layer of the circular muscle (cm). Between the muscle cells, elongated cells show NOSIP immunoreactivity (arrows). Ganglionic sheath cells (doubled arrowheads) and neuronal nuclei of the myenteric plexus (arrowheads) of the myenteric plexus displayed NOSIP immunoreactivity. (C, D) Large intestine. The nuclei and plasma membrane of enterocytes contained NOSIP immunoreactivity. (D) The muscularis mucosae (mm) and the longitudinal muscle layer (lm) were NOSIP immunoreactive. Occasionally NOSIP immunoreactivity was found in muscle cells of the innermost layer of the circular muscle (cm). Elongated cells (arrows) between the muscle cells of the circular muscle layer showed NOSIP immunoreactivity. (Bars: A, C = 20 μm; B, D = 50 μm).
Gastroenterology  , DOI: ( /gast )
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5. Fig. 4
Large intestine. Elongated NOSIP-immunoreactive cells in the muscle layer were identified as interstitial cells of Cajal by double-labeling with an antiserum against c-kit. (Bar = 10 μm).
Gastroenterology  , DOI: ( /gast )
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6. Fig. 5
(A, B) Liver. Hepatocytes showed intense nuclear concomitant with weak or absent cytoplasmic NOSIP immunoreactivity (dominating in A), or vice versa (dominating in B). Small arrows mark individual NOSIP-immunoreactive endothelial cells of a central vein. (C, D) Pancreas. Vascular smooth muscle cells (arrows in C) and nuclei of both exocrine and endocrine cells (D) were immunoreactive. iL, islet of Langerhans. (Bars = 50 μm).
Gastroenterology  , DOI: ( /gast )
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7. Fig. 6
Coimmunoprecipitation. With Western Blot, eNOS was detected in immunoprecipitates from rat small intestine homogenates by using the NOSIP antiserum. No eNOS was found by using the corresponding preimmune serum for immunoprecipitation.
Gastroenterology  , DOI: ( /gast )
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8. Fig. 7
Double-labeling immunofluorescence for NOSIP and nNOS in the myenteric plexus and tunica muscularis of the jejunum. Nuclear NOSIP immunoreactivity was seen both in nNOS-positive neurons (top) and in nNOS-negative neurons (middle). In the muscle layer, nNOS-immunoreactive axons ran close to elongated NOSIP-immunoreactive cells, but these immunoreactivities were not colocalized (bottom). (Bars = 10 μm).
Gastroenterology  , DOI: ( /gast )
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9. Fig. 8
Immunolabeling for eNOS using an antiserum raised against the peptide sequence 599–613 of eNOS. (A, A') Esophagus. Labeling of striated muscle and suprabasal epithelial cells (A) was absent after antibody preabsorption with the corresponding peptide (A'); mm, lamina muscularis mucosae. (B) Stomach, tunica muscularis. Granular and faint membrane-bound eNOS immunoreactivity was seen at the smooth muscle cells. The inset shows a myenteric ganglion with numerous eNOS-immunoreactive neurons (arrowhead) and 2 nonreactive neurons (arrow). (C, D) Large intestine. As in the stomach, the smooth muscle cells of the tunica muscularis (C) showed moderate granular and membrane-bound immunoreactivity; the arrow points to the endothelia of a longitudinally running capillary. (D) Colonic enterocytes display granular eNOS immunolabeling directly apical to the nucleus (arrow), whose position can be better resolved when (D') the same microscopical field is viewed under Nomarski optics. (E) Liver. Hepatocytes contain granular eNOS immunolabeling. (F, F') Pancreas. Acinar cells showed granular eNOS immunoreactivity (arrows). Arrowheads point to capillaries. (F') Preabsorption control. (Bars: A–C, F, F' = 20 μm; D–E = 10 μm).
Gastroenterology  , DOI: ( /gast )
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10. Fig. 9
NADPH-diaphorase reaction. (A) Esophagus. Striated muscle and epithelial cells are NADPH-diaphorase reactive. (B, C) Stomach. (B) A myenteric ganglion is located between NADPH-diaphorase–positive smooth muscle cells of the longitudinally cut circular layer (top) and the cross-sectioned longitudinal layer (bottom). Within the ganglion, 2 moderately reactive neuronal cell bodies with visible, clear nuclei are flanked by 2 intensely reactive neurons in which the nucleus is covered by reaction product. (C) Two NADPH-diaphorase–positive parietal cells of the gastric mucosa are shown at high magnification. (D, E) Large intestine. A positive NADPH-diaphorase reaction was found in the vascular smooth muscle of a subserosal artery (D) and in colonic enterocytes (E). (Bars: A, D = 20 μm; B, C, E = 10 μm).
Gastroenterology  , DOI: ( /gast )
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