1. Volume 121, Issue 4, Pages 757-766 (October 2001)
Neural stem cells express RET, produce nitric oxide, and survive transplantation in the gastrointestinal tract 
Maria–Adelaide Micci, Randall D. Learish, Hui Li, Bincy P. Abraham, Pankaj Jay Pasricha 
Gastroenterology 
Volume 121, Issue 4, Pages (October 2001)
DOI: /gast
Copyright © 2001 American Gastroenterological Association Terms and Conditions

2. Fig. 1
CNS-NSC express nNOS and produce NO in vitro. (A) Western blot analysis of total rat pituitary lysate (lane 1, as positive control) and rat CNS-NSC total proteins extract probed with (lane 2) or without (lane 3, as negative control) a specific anti-nNOS antibody. (B) nNOS immunoreactivity in rat CNS-NSC in culture for 24 hours. (C) NO production in CNS-NSC. The relative fluorescence intensity reflecting NO production by CNS NSC was measured in the presence of L-arginine (1 mmol/L; n = 11 cells) or L-NAME (100 μmol/L; n = 8 cells) at 1, 2, 3, and 4 minutes after addition of ionomycin (1.4 μmol/L). A significant increase in fluorescence intensity is observed in the presence of L-arginine as compared with L-NAME. Data are from one experiment but are representative of 3 others. Error bars indicate SD. * P < 0.05; ** P < 0.01.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Expression of nNOS in differentiated rat CNS-NSC in vitro. (A–C) Photomicrographs of cultured rat CNS-NSC double-stained for (A) βIII-tubulin, (B) nNOS, and (C) superimposition of both. (D–E) Photomicrographs of cultured rat CNS-NSC double-stained for (A) PGP 9.5, (B) nNOS, and (C) super-imposed of both. Note that nNOS immunoreactivity is present in all the βIII-tubulin and PGP 9.5-positive cells, indicating that nNOS expression is maintained in differentiated neurons. (G–I) Photomicrographs of cultured rat CNS-NSC double-stained for (A) GFAP, (B) nNOS, and (C) superimposition of both. nNOS immunoreactivity is absent in GFAP-positive cells (arrows). Calibration bars: 20 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

4. Fig. 3
CNS-NSC express the receptor complex for GDNF. (A) Western blot analysis of total proteins extracted from rat brain (lane 1, as positive control) and rat CNS-NSC probed with (lane 2) or without (lane 3, as negative control) a specific anti-RET antibody. (B) RET immunoreactivity in cultured rat CNS-NSC. Specific immunoreactivity is observed in a subpopulation of cells. (C) Western blot analysis of total proteins extracted from rat brain (lane 1, as positive control) and rat CNS-NSC with (lane 2) or without (lane 3, as negative control) anti-GFRα1 antibody. (D) GFRα1 immunoreactivity in cultured rat CNS-NSC. Panels B and D show superimposition of fluorescent and bright field images.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

5. Fig. 4
RET immunoreactivity in rat CNS-NSC that were treated for 10 days with either (A) NB27 media plus 20 ng/mL EGF and 20 ng/mL bFGF or (B) the same media plus 100 ng/mL GDNF. Cell nuclei are counterstained in blue with DAPI. Calibration bars: 20 μm. (C) The histogram demonstrates the significant increase in the percentage of RET-positive cells under the effects of 100 ng/mL GDNF as compared with control media (control). Error bars represent SE. *P = Total number of cells: n = 2350 (control); n = 2417 (GDNF) pooled from 3 independent experiments.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

6. Fig. 4
RET immunoreactivity in rat CNS-NSC that were treated for 10 days with either (A) NB27 media plus 20 ng/mL EGF and 20 ng/mL bFGF or (B) the same media plus 100 ng/mL GDNF. Cell nuclei are counterstained in blue with DAPI. Calibration bars: 20 μm. (C) The histogram demonstrates the significant increase in the percentage of RET-positive cells under the effects of 100 ng/mL GDNF as compared with control media (control). Error bars represent SE. *P = Total number of cells: n = 2350 (control); n = 2417 (GDNF) pooled from 3 independent experiments.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

7. Fig. 5
(A) DiI-labeled CNS-NSC 8 weeks after transplantation into the pyloric wall of C57BL/6J mice. Cross sections of mouse pylorus were counterstained in blue with DAPI to identify cell nuclei. To help identify the location of the grafted cells, bright field images (not shown) were superimposed with fluorescent images. DiI-labeled cells are recovered in the mouse pyloric wall where they are mainly localized between the longitudinal (LM) and circular (CM) muscle layer. (B-C) Double immunofluorescence staining of grafted DiI-labeled CNS-NSC shows colocalization of DiI with nNOS and the neuronal markers βIII-tubulin (B) and PGP 9.5 (C), indicating that at least a subpopulation of grafted cells (arrows) differentiate into neurons and maintain the expression of the enzyme nNOS. Calibration bars: 10μm. LM, longitudinal muscle; CM, circular muscle.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions