Journal:

Article Title: T Cell Apoptosis in Human Heart Allografts

doi:

Figure Lengend Snippet: Production of granzyme B and perforin during rejection. A: Granzyme B is detected in part of the lymphocytes during rejection (brown staining). Original magnification, ×150. B: Perforin is produced to the same extent as granzyme B, in part of the infiltrating cells (brown staining). Original magnification, ×150. Detection in paraffin-embedded tissue was by biotinylated horse anti-mouse antibody and HRP-conjugated streptavidin, developed with DAB.

Article Snippet: For CLSM, sections were incubated with FITC-conjugated streptavidin (1:20; lot 14594222-09, Boehringer Mannheim) for 30 minutes at RT.

Techniques: Staining, Produced

Journal:

Article Title: T Cell Apoptosis in Human Heart Allografts

doi:

Figure Lengend Snippet: Apoptosis, demonstrated by DNA-ladder analysis and nuclear condensation. A: DNA fragmentation analysis of heart tissue during allograft rejection. DNA was isolated from heart tissue during moderate to severe rejection and from healthy heart tissue. After electrophoresis on an agarose gel, the DNA ladder, characteristic for apoptosis, was observed in hearts with a rejection, but not in normal, non-transplanted heart tissue. Lanes 1 and 2, rejection grade 3A; lane 3, rejection grade 3B; lane 4, healthy heart; lane 5, DNA marker (φX 174). B: Nuclear condensation, one of the morphological characteristics of apoptosis, was observed in apoptotic cells. In situ end labeling of the DNA (TUNEL) demonstrates apoptotic cells (left panel, arrows) . Propidium iodide counterstaining of the nuclei (right panel) shows chromatin condensation in the TUNEL-positive cells (arrows) and a normal chromatin distribution in the non-apoptotic cells. Original magnification, ×550. For biotin-dUTP-labeled DNA (TUNEL), detection was by TRITC-conjugated streptavidin.

Article Snippet: For CLSM, sections were incubated with FITC-conjugated streptavidin (1:20; lot 14594222-09, Boehringer Mannheim) for 30 minutes at RT.

Techniques: Isolation, Electrophoresis, Agarose Gel Electrophoresis, Marker, In Situ, End Labeling, TUNEL Assay, Labeling

Journal:

Article Title: T Cell Apoptosis in Human Heart Allografts

doi:

Figure Lengend Snippet: Regulation of apoptosis in EMBs during heart allograft rejection. A: FasL expression on apoptotic cells. Double staining for FasL (green) and TUNEL (red) demonstrated that virtually all apoptotic cells expressed FasL. Original magnification, ×280. B: FasL expression on CD4+ T cells. The majority of CD4+ T cells (green) co-expressed FasL (red, arrowheads). Original magnification, ×210. C: FasL expression on macrophages. All CD68+ macrophages (green) co-expressed FasL (red). Original magnification, ×190. D: Fas expression, shown by conventional immunoperoxidase staining (brown). Most of the infiltrating cells expressed Fas. Original magnification, ×100. E: Bax expression in apoptotic cells. Bax (green) was expressed on 50% to 70% of all apoptotic cells (TUNEL, red). Original magnification, ×330. F: Bcl2 expression in the infiltrate. Approximately 50% of all cells expressed Bcl2 (brown staining). Original magnification, ×250. For FasL, detection was by SWARPO and direct (green) or indirect (red) signal amplification. For TUNEL, detection was by TRITC-conjugated streptavidin. For CD4 and CD68, staining was with FITC-conjugated antibodies and direct signal amplification. For Bax, detection was by RAMPO and direct signal amplification. For Fas and Bcl2, detection was by biotinylated horse anti-mouse antibody and HRP-conjugated streptavidin, developed with DAB.

Article Snippet: For CLSM, sections were incubated with FITC-conjugated streptavidin (1:20; lot 14594222-09, Boehringer Mannheim) for 30 minutes at RT.

Techniques: Expressing, Double Staining, TUNEL Assay, Immunoperoxidase Staining, Staining, Amplification