1. Volume 129, Issue 6, Pages 2047-2057 (December 2005)
erbB-2/neu Transformed Rat Cholangiocytes Recapitulate Key Cellular and Molecular Features of Human Bile Duct Cancer 
Guan–Hua Lai, Zichen Zhang, Xue–Ning Shen, Deanna J. Ward, Jennifer L. DeWitt, Shawn E. Holt, Rebecca A. Rozich, Douglas C. Hixson, Alphonse E. Sirica 
Gastroenterology 
Volume 129, Issue 6, Pages (December 2005)
DOI: /j.gastro
Copyright © 2005 American Gastroenterological Association Terms and Conditions

2. Figure 1
(A) DNA sequence analysis of genomic DNA from rat BDE1 cells infected retrovirally with Glu664neu (BDEneu), showing a transversion point mutation at nucleotide 2012 (arrow) corresponding to that determined for (B) plasmid pJRneu carrying the erbB-2/neu gene showing the same mutation at the nucleotide 2012 hot spot (positive control). (C) Corresponding DNA sequence of wild-type rat erbB-2/neu gene carried in plasmid pSVneu (nonmutated control).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

3. Figure 2
(A) Karyotype analysis of cultured parent BDE1 cells compared with that of (B) the BDEneu cell line after retroviral infection with Glu664neu and G418 selection. For each cell line, chromosome counts were made on a total of 50 metaphase spreads.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

4. Figure 3
(A) Representative phase-contrast photomicrograph showing the epithelial nature of BDEneu cells cultured to confluence on a plastic substratum. (B) Phase-contrast photomicrograph showing typical anchorage-independent colonies of BDEneu cells formed in soft agar at 2 weeks after initial cell seeding of 1500 cells. (C) Comparative anchorage-dependent growth curves for BDEneu cells vs parent BDE1 cells cultured on rat-tail type 1 collagen-coated plastic wells. Each value represents the mean ± SD from individual cell counts made on triplicate cultures per time point. Cell population doubling time: BDEneu = 18.9 hours; BDE1 = 21.3 hours. (D) Differences in the number of cell colonies formed after 2 weeks of anchorage-independent growth of BDEneu cells in soft agar compared with that of BDE1 cells. Each value represents the mean ± SD from cell colony counts made on 3 separate cultures per cell line. *P ≤ .001.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

5. Figure 4
(A) Representative telomeric repeat amplification protocol assay showing markedly increased telomerase activity expressed by cultured BDEneu cells compared with that of the BDEneo and BDE1 cell lines. Note that the baseline Q value (see Materials and Methods section for explanation of Q value) determined for BDEneo control cells was unchanged from that of parent BDE1 cells. The C611B ChC rat cholangiocarcinoma cell line served as a positive control. (B) PGE2 overproduction from arachidonic acid by cultured BDEneu cells compared with PGE2 elaborated into the medium by comparably maintained BDE1 and BDEneo control cells. Each value represents the mean ± SD from individual measurements made on 3 separate cultures per cell line. P ≤ .001 compared with BDEneo and BDE1.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

6. Figure 5
Representative Western blot analysis showing relevant altered protein expression and phosphorylation profiles of BDEneu cells compared with those of the BDE1 and BDEneo control cell lines. ripa, no-cell buffer (negative) control. The mean protein-band density ratios reflect the mean band intensity for each protein of interest, as determined by densitometry, relative to that of corresponding β-actin, which served as the internal housekeeping standard.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

7. Figure 6
Representative real-time RT-PCR amplification plots for (A) ErbB-2/Neu, (B) COX-2, and (C) MUC1 apoprotein mRNA levels compared with glyceraldehyde-3-phosphate dehydrogenase mRNA expressed in cultured BDEneu cells relative to corresponding mRNA levels for these gene products expressed in BDEneo control cells.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

8. Figure 7
(A) Gross pathology of an invasive tumor formed in the liver of a Fischer 344 rat at 1 month after orthotopic cell transplantation of 4 × 106 BDEneu cells inoculated via the left hepatic bile duct compared with control liver transplanted with BDEneo cells. Arrows point to left liver lobe, which was bisected to expose the intrahepatic tumor. Note the irregular margins of the mass-forming tumor, which has replaced much of the normal liver tissue of the left hepatic and median lobes. The right liver lobe (rt) did not show evidence of tumor, but the hepatic hilus (*) at the level of the left hepatic bile duct was replaced by tumor. (B) No tumor was detected in liver comparably transplanted with BDEneo cells. (B–E) Representative photomicrographs showing strong uniform positive immunoreactivity of neoplastic ducts for biliary CK19, plasma membrane ErbB-2/Neu, ErbB-2/Neu immunoreactive for phosphotyrosine1248, and cytoplasmic COX-2, respectively, in desmoplastic ductal carcinoma developed in rats transplanted with BDEneu cells. (F) The low level of mucicarmine histochemical staining of mucin (red) present in neoplastic ducts of ductal carcinoma originated from transplanted BDEneu cells.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions