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Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. <t>Tubulin</t> levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.
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Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. <t>Tubulin</t> levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.
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Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. <t>Tubulin</t> levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.
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Jackson Immuno sheep anti mouse igg conjugated to hrp
Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. <t>Tubulin</t> levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.
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Image Search Results


Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. Tubulin levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.

Journal: The American Journal of Pathology

Article Title: Regulation of Apo2L/Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand-Induced Apoptosis in Thyroid Carcinoma Cells

doi: 10.1016/s0002-9440(10)64220-4

Figure Lengend Snippet: Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. Tubulin levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.

Article Snippet: Recombinant human TRAIL was obtained from Immunex Corporation (Seattle, WA), in a leucine zipper (LZ) form that promotes and stabilizes the formation of trimers, as previously reported;13 for comparison, several experiments were repeated using the recombinant Apo2L form from Genentech Inc. (South San Francisco, CA).20 The goat polyclonal antibodies for DR4, DR5, DcR1, and mouse monoclonal antibody for tubulin were from Santa Cruz Biotechnologies (Santa Cruz, CA); the anti-DcR2 rabbit polyclonal antibody was from Imgenex (San Diego, CA); cycloheximide, geldanamycin, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was from Sigma Chemical Co. (St Louis, MO); IGF-1, bFGF, EGF, IFN- , and TNF- were from R&D Systems (Minneapolis, MN; bisindolylmaleimide (BIM) III and wortmannin were from Calbiochem (La Jolla, CA); rabbit polyclonal antibody for caspase-10 was from Research Diagnostics Inc. (Flanders, NJ); the IGF-1 receptor neutralizing antibody aIR3 was from Oncogene Research (Cambridge, MA); and the enhanced chemiluminescence (ECL) kit, which includes the peroxidase-labeled anti-mouse and anti-rabbit secondary antibodies, was from Amersham (Arlington Heights, IL).

Techniques: MTT Assay, Comparison, Control