TRAIL-R deficiency in mice enhances lymph node metastasis without affecting primary tumor development
J. Clin. Invest. Anne Grosse-Wilde, et al. 118:100 doi:10.1172/JCI33061 [
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Figure 4Detachment sensitizes skin carcinoma cells specifically to TRAIL and CD95L.
(
A) Subconfluent cells were incubated in the absence (control) or presence of TRAIL (iz-muTRAIL, 1 μg/ml) and/or bortezomib (20 nM) for 24 hours. Cell death was determined by PI staining and flow cytometric analysis. (
B and
C) Cells were trypsinized and cultured either on an uncoated (attached) or a poly-HEMA–coated surface (detached) in the absence or presence of TRAIL (
B; 1 μg/ml) or at the indicated concentrations (
C). (
D) Subconfluent cells (DT02) were treated with or without 1 μM thapsigargin (Tgg), 30 μg/ml oxaliplatin (Oxp), 5 μg/ml cycloheximide (Chx), or 43°C heat shock (HS) or irradiated with 15 Gy (IR) and were incubated in the absence or presence of TRAIL (1 μg/ml) for 24 hours. (
E) Attached (att) or detached (det) cells were incubated in the absence or presence of TRAIL (1 μg/ml) or CD95L (100 ng/ml) for 24 hours. Each data point represents one of the cell lines DT01–DT06 or DT11. The mean percentage of dead WT cells is indicated by a horizontal line. (
F) Attached or detached cells were incubated in the absence or presence of TRAIL (1 μg/ml), CD95L (100 ng/ml), 5-FU (200 μg/ml), or etoposide (10 μM) for 24 hours. Standard deviation for duplicates or triplicates is shown. Results are representative of at least 2 independent experiments.