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. 2007 Dec 13;118(1):100–110. doi: 10.1172/JCI33061

Figure 4. Detachment sensitizes skin carcinoma cells specifically to TRAIL and CD95L.

Figure 4

(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.