1. In vivo imaging of S-TRAIL-mediated tumor regression and apoptosis
Khalid Shah, Ching-Hsuan Tung, Xandra O. Breakefield, Ralph Weissleder 
Molecular Therapy 
Volume 11, Issue 6, Pages (June 2005)
DOI: /j.ymthe
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

2. Fig. 1
S-TRAIL-induced tumor regression in vivo. (A) Gli36fluc+ cells were infected with S-TRAIL or HGCX control vector at an m.o.i. of 1 for 2 h at 37°C and mice were implanted with either a mix of 1.5 × 106 S-TRAIL amplicon-infected and 6.0 × 106 noninfected cells (dashed circle around the tumor) or a mix of the same number of control HGCX infected and noninfected cells in the flanks. Mice bearing subcutaneous Gli36fluc+ gliomas were injected ip with d-luciferin and imaged for Fluc activity. Two representative mice are shown and each image represents a scan time of 1 min, and the pseudocolor image represents the spatial distribution of photon counts produced by active luciferase within the tumor. (B) Fluc bioluminescence intensity of tumors over time given as an average of tumors in 20 animals, with error bars representing mean ± 95% confidence intervals.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

3. Fig. 2
Imaging apoptosis in culture. (A) S-TRAIL dose response. Gli36fluc+ cells were infected with different m.o.i. of S-TRAIL amplicon and 14 h later, DEVD–luciferin compound was added and the cells were imaged for apoptosis by Fluc imaging. (B) Camptothecin dose response. Gli36fluc+ cells were treated with different concentrations of camptothecin for 6 h and imaged for apoptosis by Fluc imaging as above. Each point represents the average of three wells. (C) Substrate time curves. Gli36fluc+ cells were incubated with d-luciferin and camptothecin-treated Gli36fluc+ cells were incubated with DEVD–luciferin and both were imaged at different time points for 5 h.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

4. Fig. 3
Dual-luciferase imaging of tumor cell progression and apoptosis in culture. (A and B) Gli36fluc+ glioma cells were either infected with S-TRAIL or treated with camptothecin and imaged for cell number after incubation with luciferin. (C and D) Cells used in (A) were washed and 4 h later were incubated with DEVD–luciferin and imaged for apoptosis. Each point represents the average of five wells. (E–G) Twelve hours after infection or treatment of Gli36fluc+ cells with camptothecin, immunohistochemistry with anti-cleaved-caspase-3 antibodies was performed: (E) camptothecin-treated cells, (F) S-TRAIL vector-infected cells, and (G) control vector-infected cells.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

5. Fig. 4
Dual-luciferase imaging in vivo. Mice were implanted with a mix of S-TRAIL-infected and noninfected Gli36fluc+ cells (dotted circle) or control vector-infected and noninfected Gli36fluc+ cells in the flanks. (A) A cartoon of the implantation sites in mice is shown. (B) Two days later mice were imaged for Fluc activity and tumor progression by injecting luciferin. (C) Twenty-four hours later, mice in B were injected with DEVD-luciferin and imaged for apoptosis by Fluc imaging. One representative mouse is shown and the image represents a scan time of 1 min. Mice were sacrificed on day 3 after tumor cell implantation, and tumors were sectioned and stained by immunohistochemistry with anti-caspase-3 antibodies. The stained sections were counterstained with hematoxylin. (D) S-TRAIL-expressing tumor sections. Caspase-3-stained cells are shown by arrows. (E) Control vector expressing tumor sections (10× original magnification).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions