Journal: Gene therapy

Article Title: Angiogenesis induced in muscle by a recombinant adenovirus expressing functional isoforms of basic fibroblast growth factor.

doi: 10.1038/sj.gt.3300950

Figure Lengend Snippet: Figure 2 Test of the recombinant bFGF protein bioactivity. The data rep- resent means ± s.d. of surviving BALB/c-3T3 fibroblasts counted in tripli- cate wells. (a) 293 Cell extracts were mixed in three-fold dilutions (1/4 to 1/354) with serum-free medium. Grey bars: non-infected cells, white bars: Ad-RSVbgal-infected cells, black bars: Ad-RSVbFGF-infected cells. (b) Specific neutralization of the BALB/c-3T3 survival effect with a poly- clonal anti-bFGF antibody. C2 cell extracts (diluted 1/200 in serum-free medium) were preincubated with different concentrations (100, 30, 10, 3 and 1 mg/ml) of either an anti-bFGF polyclonal antibody (black bars) or an irrelevant antibody of the same isotype (white bars). Positive controls and negative controls are cells in 20% serum and serum-free medium, respectively.

Article Snippet: Pathology and immunohistochemistry on skeletal muscle bFGF immunodetection: Sections of gastrocnemius muscle were immunostained with different polyclonal antibodies directed against bovine bFGF (AB-33-NA; R&D Systems), E. coli-derived human bFGF (AB-233-NA), or E. coli b-galactosidase (ICN Pharmaceuticals, USA), respectively, using an immunoperoxidase protocol.46 Deparaffined 5 mm thick sections were incubated in blocking buffer (0.5% Triton X100, 3% (w/v) milk in PBS) for 30 min at RT.

Techniques: Recombinant, Infection, Neutralization

Journal: Gene therapy

Article Title: Angiogenesis induced in muscle by a recombinant adenovirus expressing functional isoforms of basic fibroblast growth factor.

doi: 10.1038/sj.gt.3300950

Figure Lengend Snippet: Figure 5 b-Galactosidase and bFGF immunostaining on muscle sections. Muscles injected with Ad-RSVbFGF (a, b) or the contralateral muscle injected with Ad-RSVbgal (c, d) were immunostained with different antibodies recognizing bFGF (a–c) or E. coli b-galactosidase (d). Anti-bFGF antibody AB- 33-NA (R&D Systems) was used in (a) and (c). It showed both an intracellular and an extracellular matrix staining (a). Anti-bFGF antibody AB-233- NA (R&D Systems) detected mostly nuclear-located bFGF isoforms (b). No bFGF staining was observed in (c), but a nuclear b-galactosidase staining was detected in the adjacent section (d). The stars point out the same location in (c) and (d).

Article Snippet: Pathology and immunohistochemistry on skeletal muscle bFGF immunodetection: Sections of gastrocnemius muscle were immunostained with different polyclonal antibodies directed against bovine bFGF (AB-33-NA; R&D Systems), E. coli-derived human bFGF (AB-233-NA), or E. coli b-galactosidase (ICN Pharmaceuticals, USA), respectively, using an immunoperoxidase protocol.46 Deparaffined 5 mm thick sections were incubated in blocking buffer (0.5% Triton X100, 3% (w/v) milk in PBS) for 30 min at RT.

Techniques: Immunostaining, Muscles, Injection, Staining