1. Isoforms and splice variant of transforming growth factor β–binding protein in rat hepatic stellate cells 
Wenrong Gong, Sylke Roth, Kristin Michel, Axel M. Gressner 
Gastroenterology 
Volume 114, Issue 2, Pages (February 1998)
DOI: /S (98)
Copyright © 1998 American Gastroenterological Association Terms and Conditions

2. Fig. 1
APAAP immunostainings of LTBP, LAP, and TGF-β in rat HSCs cultured in DMEM supplemented with 0.2% fetal calf serum for 48 hours. A polyclonal panspecific antibody against (A) TGF-β1, 2, and 3, (B) a monoclonal antibody against LAP, and (C) a polyclonal antibody against LTBP (ab 39) were used. Negative controls were performed by using nonspecific rabbit IgG instead of the specific antibodies (original magnification 400×).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Detection of the subcellular localization of LTBP, LAP, and TGF-β in MFBs by immunofluorescence staining. Cells were kept for 3 days in DMEM and 0.2% fetal calf serum after initiating the subculture of HSCs. A polyclonal antibody against (A) TGF-β1, 2, and 3; (B and D) a monoclonal antibody against LAP; and (C and E) a polyclonal antibody against LTBP (ab 39) were used. (E) LTBP staining was compared with (F) the localization of talin, which was detected by a monoclonal antibody against talin. Negative controls were performed by using nonspecific rabbit or mouse IgG instead of the specific antibodies (G and H) (original magnification 200×).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Expression of the LTBP isoforms in HSCs, MFBs, and various rat tissues. RT-PCR of total RNA from HSCs cultured for the indicated times and MFBs cultured for 3 days was used to determine the expression of (A) LTBP-1, (B) LTBP-2, and (C) LTBP-3. (D) The expression pattern of LTBP-1 in various rat tissues (H, heart; B, brain; Li, liver; K, kidney; Lu, lung; S, spleen) and of LTBP-2 and LTBP-3 in rat liver. All negative controls were performed without reverse transcription (NC). The numbers on the right give the size of some molecular mass markers.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

5. Fig. 4
cDNA sequence of LTBP isoforms and structure of large latent TGF-β complex. (A) cDNA sequence alignment of LTBP-1D (upper lane) and LTBP-1 (middle lane). LTBP-1D shows an inframe deletion of 159 bases. The deduced amino acid is listed in the lower lane and the spliced sequence is indicated in boldface. The postulated endopeptidase cleavage site is underlined. (B) Part of identified rat LTBP-2 and -3 cDNA sequences corresponding to the hinge region of LTBP-1. (C) The structure of large latent TGF-β complex with presentation of yet known LTBP-1 splice variants. Schematic model described by Saharinen et al.26 shows the extracellular matrix–associated large latent TGF-β complex containing LTBP-1, the covalent bound LAP dimer that associates noncovalently the TGF-β dimer. Smaller and larger LTBP-1 splice variants differ in their N-terminal part, which influences the association to extracellular matrix.25 The identified deletion of LTBP-1D in the hinge region was marked.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

6. Fig. 4
cDNA sequence of LTBP isoforms and structure of large latent TGF-β complex. (A) cDNA sequence alignment of LTBP-1D (upper lane) and LTBP-1 (middle lane). LTBP-1D shows an inframe deletion of 159 bases. The deduced amino acid is listed in the lower lane and the spliced sequence is indicated in boldface. The postulated endopeptidase cleavage site is underlined. (B) Part of identified rat LTBP-2 and -3 cDNA sequences corresponding to the hinge region of LTBP-1. (C) The structure of large latent TGF-β complex with presentation of yet known LTBP-1 splice variants. Schematic model described by Saharinen et al.26 shows the extracellular matrix–associated large latent TGF-β complex containing LTBP-1, the covalent bound LAP dimer that associates noncovalently the TGF-β dimer. Smaller and larger LTBP-1 splice variants differ in their N-terminal part, which influences the association to extracellular matrix.25 The identified deletion of LTBP-1D in the hinge region was marked.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

7. Fig. 5
Secretion of LTBP from metabolically labeled rat HSCs and MFBs. Cells were labeled with [35S]cysteine and [35S]methionine in cysteine- and methionine-free medium for 6 hours at 37°C. The cell-conditioned medium was then subjected to immunoprecipitation using anti-LTBP (ab 39). Negative control was performed using a nonimmune rabbit serum (NC). Samples were analyzed by SDS–gel electrophoresis (3%–12% polyacrylamide) under nonreducing and reducing conditions followed by fluorography. Positions of molecular mass standards (kilodaltons) are indicated. The numbered proteins (1–5) are discussed in the text.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

8. Fig. 6
Expression of the TGF-β isoforms in rat HSCs and MFBs. (A) TGF-β1, (B) TGF-β2, and (C) TGF-β3 were determined by RT-PCR of total RNA from HSCs cultured for the indicated times and MFBs cultured for 3 days. The negative controls were performed without reverse transcription (NC). Some molecular mass markers are indicated.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

9. Fig. 7
APAAP immunostainings of TGF-β isoforms in rat HSCs (left panel) and MFBs (right panel) cultured for 48 hours. Polyclonal antibodies against (A and B) TGF-β1, (C and D) TGF-β2, and (E and F) TGF-β3 were used. Negative controls were performed by using nonspecific rabbit IgG instead of the specific antibodies (G and H) (original magnification 200×).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions