Journal: BMC Cancer

Article Title: Combination therapy with c-met inhibitor and TRAIL enhances apoptosis in dedifferentiated liposarcoma patient-derived cells

doi: 10.1186/s12885-019-5713-2

Figure Lengend Snippet: The c-Met inhibitor PF enhanced TRAIL-mediated apoptosis in liposarcoma. PF enhances TRAIL-mediated apoptosis in DDLPS cell lines and PDCs. Shown are the cell viabilities of the established cell lines ( a ) LPS224 and LPS246 and the PDCs ( b ) 11GS-013 and 11GS-079 after 48 h of incubation with 5 μM PF and 5 ng/ml rhTRAIL under the following treatment schemes: negative control, PF alone for 48 h; rhTRAIL alone for 48 h; PF for 24 h followed by rhTRAIL for 24 h; rhTRAIL for 24 h followed by PF for 24 h; and concurrent treatment with PF and rhTRAIL for 48 h. We analyzed apoptosis using annexin V and 7-AAD ( c and d )

Article Snippet: Human recombinant TRAIL protein (rhTRAIL) was purchased from R&D Systems (R&D Systems, Minneapolis, MN, USA). c-Met inhibitors (PHA665752: PHA; PF02341066:crizotinib: PF) were bought from Sellekchem (Houston, TX, USA).

Techniques: Incubation, Negative Control

Journal: BMC Cancer

Article Title: Combination therapy with c-met inhibitor and TRAIL enhances apoptosis in dedifferentiated liposarcoma patient-derived cells

doi: 10.1186/s12885-019-5713-2

Figure Lengend Snippet: Efficacy of treatment with rhTRAIL in sarcoma cell lines. Cell viability of ADMSCs ( a ), MFH-ino ( b ), SW872 ( c ), and HT1080 ( d ) after 48 h of incubation with serial dilutions of rhTRAIL protein (0–10 ng/ml)

Article Snippet: Human recombinant TRAIL protein (rhTRAIL) was purchased from R&D Systems (R&D Systems, Minneapolis, MN, USA). c-Met inhibitors (PHA665752: PHA; PF02341066:crizotinib: PF) were bought from Sellekchem (Houston, TX, USA).

Techniques: Incubation

Journal: BMC Cancer

Article Title: Combination therapy with c-met inhibitor and TRAIL enhances apoptosis in dedifferentiated liposarcoma patient-derived cells

doi: 10.1186/s12885-019-5713-2

Figure Lengend Snippet: Efficacy of tumor cell suppression through combined treatment with the c-Met inhibitor PF and rhTRAIL in DDLPS PDCs. Combination treatment with PF and rhTRAIL suppressed cell viability effectively in the DDLPS established cell lines: LPS246 ( a ) and LPS224 ( b ); and in the DDLPS PDCs: 11GS-013 ( c ), 11GS-079 ( d ), 11GS-099 ( e ), 11GS-106 ( f ), 14GS-026 ( g ), and 14GS-076 ( h )

Article Snippet: Human recombinant TRAIL protein (rhTRAIL) was purchased from R&D Systems (R&D Systems, Minneapolis, MN, USA). c-Met inhibitors (PHA665752: PHA; PF02341066:crizotinib: PF) were bought from Sellekchem (Houston, TX, USA).

Techniques:

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 1. rhTRAIL exposure effect alone and in combination with WT HSV-1 virus on HNSCC cell lines and the normal cell line, HaCaT. Treatment was performed with rhTRAIL at 100 ng/mL and in combination with WT HSV-1 virus at MOI 0f. 0.2 and 1.0. Cell viability analysis after 48 (A) and 72 h (B) of exposure. Results are expressed by mean percentage ± standard deviation of cell viability relative to PBS 1X (considering 100% viability). Data represent the average of three independent assays performed in triplicate. The asterisks indicate statistical significance (*P < 0.05; **P < 0.01; ***P < 0.001) in the comparison with the experimental groups by non-normal distribution Kruskal–Wallis test.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Virus, Standard Deviation, Comparison

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 2. Expression analysis of apoptosis-associated proteins in the HCB289 cell line after 24 h of exposure with rhTRAIL ligand, WT HSV-1 and in combination. (A) and (B) detection and quantification of proteins after treatment. Results are expressed as mean percentage ± standard deviation of relative expression of cleaved protein relative to control (considering 100% expression) and normalized by the portion of total protein (non- cleaved). Data represent the average of three independent assays. The asterisks indicate statistical significance (*P < 0.05; **P < 0.01; ***P < 0.001) in the comparison with the experimental groups by one-way ANOVA. Raw blots are presented in Supplementary Data 1.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Expressing, Standard Deviation, Control, Comparison

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 3. Expression analysis of apoptosis-associated proteins in the UD-SCC-2 cell line after 24 h of exposure with rhTRAIL ligand, WT HSV-1 and in combination. (A) and (B) detection and quantification of proteins after treatment. Results are expressed as mean percentage ± standard deviation of relative expression of cleaved protein relative to control (considering 100% expression) and normalized by the portion of total protein (non- cleaved). Data represent the average of three independent assays. The asterisks indicate statistical significance (*P < 0.05; **P < 0.01; ***P < 0.001) in the comparison with the experimental groups by one-way ANOVA. Raw blots are presented in Supplementary Data 2.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Expressing, Standard Deviation, Control, Comparison

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 4. Expression analysis of apoptosis-associated proteins in the HCB289 cell line after 48 h of exposure with rhTRAIL ligand, WT HSV-1 and in combination. (A) and (B) detection and quantification of proteins after treatment. Results are expressed as mean percentage ± standard deviation of relative expression of cleaved protein relative to control (considering 100% expression) and normalized by the portion of total protein (non- cleaved). Data represent the average of two independent assays. The asterisks indicate statistical significance (*P < 0.05) in the comparison with the experimental groups by one-way ANOVA. Raw blots are presented in Supplementary Data 3.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Expressing, Standard Deviation, Control, Comparison

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 5. Expression analysis of apoptosis-associated proteins in the UD-SCC-2 cell line after 48 h of exposure with rhTRAIL ligand, WT HSV-1 and in combination. (A) and (B) detection and quantification of proteins after treatment. Results are expressed as mean percentage ± standard deviation of relative expression of cleaved protein relative to control (considering 100% expression) and normalized by the portion of total protein (non- cleaved). Data represent the average of two independent assays. The asterisks indicate statistical significance (*P < 0.05) in the comparison with the experimental groups by one-way ANOVA. Raw blots are presented in Supplementary Data 4.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Expressing, Standard Deviation, Control, Comparison

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 6. Flow cytometry apoptosis analysis after treatment with rhTRAIL ligand, WT HSV-1 and in combination. A and B) Analysis of HCB289 cell line after 24 and 48 h of treatment exposure. C and D) Analysis of UD-SCC-2 cell line after 24 and 48 h of treatment exposure. Results are expressed as mean percentage ± standard deviation of the cell death rate. Data represent the average of three independent assays. The asterisks indicate statistical significance (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.001) in the comparison with the experimental groups by one-way ANOVA.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Flow Cytometry, Standard Deviation, Comparison

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 7. DR-5 expression analysis in the HCB289 cell line after treatment with rhTRAIL ligand, WT-HSV-1 and combination. (A) and (B) Detection and quantification of the receptor after 24 h of exposure treatment. (C) and (D) Detection and quantification of the receptor after 48 h of exposure treatment. Results are expressed as mean percentage ± standard deviation of relative expression of cleaved protein relative to control (considering 100% expression) and normalized by the protein α-tubulin. Data represent the average of two independent. Raw blots are presented in Supplementary Data 5.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Expressing, Standard Deviation, Control

Journal: Scientific reports

Article Title: Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

doi: 10.1038/s41598-023-44888-9

Figure Lengend Snippet: Figure 8. DR-5 expression analysis in the UD-SCC-2 cell line after treatment with rhTRAIL ligand, WT-HSV-1 and combination. (A) and (B) Detection and quantification of the receptor after 24 h of exposure treatment. (C) and (D) Detection and quantification of the receptor after 48 h of exposure treatment. Results are expressed as mean percentage ± standard deviation of relative expression of cleaved protein relative to control (considering 100% expression) and normalized by the protein α-tubulin. Data represent the average of two independent assays. Raw blots are presented in Supplementary Data 6.

Article Snippet: :(0123456789) Scientific Reports | (2023) 13:18023 | https://doi.org/10.1038/s41598-023-44888-9 treated with rhTRAIL (#375-TL, R&D Systems, Minneapolis, MN, EUA) at 100 ng/mL, HSV-1 MOI 0.2 and/ or HSV-1 MOI 1.

Techniques: Expressing, Standard Deviation, Control

Journal: Cancer Gene Therapy

Article Title: Targeting GD2-positive glioblastoma by chimeric antigen receptor empowered mesenchymal progenitors

doi: 10.1038/s41417-018-0062-x

Figure Lengend Snippet: GBM target cells characterization. a Representative histograms showing GD2 expression, dark gray curve, on human T98G (97 ± 1%), U87MG (57 ± 13%), and A172 (2 ± 1%) GBM cell lines by FACS. APC-conjugated secondary Ab was used as isotype and represented by light gray line. b Expression of both agonistic (DR4 and DR5) and decoy (DcR1, DcR2) TRAIL receptors on GBM cell lines by FACS. c Sensitivity of GBM tumor cells to apoptosis induced by recombinant human TRAIL (rhTRAIL). T98G cell viability by supravital propidium iodide (PI) staining, U87MG and A172 cell viability by MTS assay after 24 h of rhTRAIL treatment at different doses in comparison with untreated control (CTR). p < .05 by Student’s t test between the highest rhTRAIL dose (1000 ng/ml) and untreated CTR, for all GBM lines. Data are expressed as mean ± SD

Article Snippet: After 12 h, different concentrations (10 ng, 50 ng, 100 ng, 500 ng, and 1000 ng/ml) of recombinant human TRAIL (rhTRAIL) (Peprotech, London, UK) were added in the culture media.

Techniques: Expressing, Recombinant, Staining, MTS Assay, Comparison, Control

Journal: Cancer Gene Therapy

Article Title: Targeting GD2-positive glioblastoma by chimeric antigen receptor empowered mesenchymal progenitors

doi: 10.1038/s41417-018-0062-x

Figure Lengend Snippet: Bi-functional MSCs exert in vitro cytotoxicity on target GBM cell lines. a In vitro impact of bi-functional MSCs against a T98G, b U87MG, c A172 GBM lines, and d primary C3c GBM cells testing multiple target-to-effector ratios (1:1, 1:2, and 1:5). Tumor cell death by supravital propidium iodide (PI) for T98G, A172, and C3c and by Annexin V/PI staining for U87MG after 24 h (left column) and 48 h (right column). Recombinant human TRAIL (rhTRAIL, 1μg/ml) was used as a positive control of cell death, while tumor cell lines alone as a negative control (CTR). Reported p values regard multiple comparisons among mTRAIL MSCs and bi-functional MSC conditions versus control groups represented by EV MSCs, GD2 tCAR MSCs, rhTRAIL, or CTR. For T98G, * p < .05, ° p < .01, § p < .01; for U87MG, * p < .05, ° p < .05, § p < .05; for A172, * p < .05, ° p < .01, § p < .05; for C3c, * p < .05, ° p < .001, § p < .00. All p values have been calculated by Student’s t test. Data are expressed as mean ± SD

Article Snippet: After 12 h, different concentrations (10 ng, 50 ng, 100 ng, 500 ng, and 1000 ng/ml) of recombinant human TRAIL (rhTRAIL) (Peprotech, London, UK) were added in the culture media.

Techniques: Functional Assay, In Vitro, Staining, Recombinant, Positive Control, Negative Control, Control

Journal: Cancer biology & therapy

Article Title: TRAIL induces endocytosis of its death receptors in MDA-MB-231 breast cancer cells.

doi: 10.4161/cbt.8.10.8141

Figure Lengend Snippet: Figure 1. TRAIL undergoes internalization in MDA-MB-231 but not BT474 breast cancer cells. (A) Cells ( 1 x 106 cells/mL) were first incubated in complete medium supplemented with or without 100 ng/mL of rhTRAIL at 4°C for 10 min, followed by incubation with 10 μg/mL of anti-TRAIL-PE for an additional 10 min. The mixtures were then incubated at 37°C for the indicated times (0–60 min). After extensive washes as described in Material and Methods, cells were resuspended in 0.5 mL cold PBS and analyzed by flow cytometry. The left panel shows a histogram for selected times during a kinetic analysis of TRAIL internalization, and the right panel shows the time-dependent increase in the mean fluorescence intensity. (B) MDA-MB-231 cells were grown on glass chamber slides, treated as in (A), fixed, and analyzed by confocal microscopy. Nuclei were visualized by Hoechst33342 staining (blue). (C) Cell surface expression of TRAIL receptors was assessed by FACS analysis after staining with PE-conjugated monoclonal antibodies to DR4, DR5, DcR1 and DcR2, respectively. Shadowed his- tograms represent control cells stained with isotype-matched control IgG. Right-shift indicates the pres- ence of receptors on cell surface. Data shown are representative of three independent experiments.

Article Snippet: Recombinant human TRAIL (rhTRAIL) expressed and purified as homotrimeric protein from E. coli, and monoclonal antibodies specific to TRAIL death receptor-4 (AF347) and -5 (MAB6312) and their phycoerythrin (PE)-conjugated forms were purchased from R&D Systems (Minneapolis, MN).

Techniques: Incubation, Flow Cytometry, Fluorescence, Confocal Microscopy, Staining, Expressing, Bioprocessing, Control

Journal: Cancer biology & therapy

Article Title: TRAIL induces endocytosis of its death receptors in MDA-MB-231 breast cancer cells.

doi: 10.4161/cbt.8.10.8141

Figure Lengend Snippet: Figure 2. TRAIL downregulates cell surface expression of DR4 and DR5 in MDA-MB-231 cells. (A) MDA-MB-231 cells were treated with TRAIL (100 ng/mL) at 37°C for 0, 10 or 30 min, and analyzed by FACS for the remaining surface DR4, DR5, Her-2, epidermal growth factor recep- tor (EGFR), respectively, as described in 1C. (B) The percent of receptors remaining on cell surface was estimated by the median fluorescence intensities at the indicated times (Ft) relative to their basal levels (F0); receptor remaining % = (Ft - FIgG)/(F0 - FIgG), where FIgG is the median fluorescence of cells stained with control IgG-PE (shaded histogram). (C) MDA-MB-231 cells were transiently transfected with a plasmid for GFP fusion protein of DR4 (GFP-DR4). After 24 h post-transfection, cells were treated with rhTRAIL (100 ng/mL) for the indicated times at 37°C, counterstained with CellTracker red, fixed and analyzed by confocal microscope.

Article Snippet: Recombinant human TRAIL (rhTRAIL) expressed and purified as homotrimeric protein from E. coli, and monoclonal antibodies specific to TRAIL death receptor-4 (AF347) and -5 (MAB6312) and their phycoerythrin (PE)-conjugated forms were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Fluorescence, Staining, Control, Transfection, Plasmid Preparation, Microscopy

Journal: Cancer biology & therapy

Article Title: TRAIL induces endocytosis of its death receptors in MDA-MB-231 breast cancer cells.

doi: 10.4161/cbt.8.10.8141

Figure Lengend Snippet: Figure 3. DR4 is cleaved after internalization. (A) TRAIL induces proteolytic cleavage of endogenous DR4, but not DR5. Cells were treated with TRAIL (20 ng/mL) for the indicated times and analyzed by west- ern blotting. RhoGDI, a caspase-resistant protein,26 was used as a loading control. Caspase activation is indicated by the cleavage of pro-caspase 8 (C-8) and caspase-3 (C-3) and the appearance of their frag- ments p43/41, p18 and p20/17. (B) The percentage of remaining intact receptor (%) and caspase activities were quantified by densitometry analysis of the bands in (A) and are expressed relative to untreated cells (Time 0). (C) Cleavage of DR4 is inhibited by caspase inhibitor. Cells were pretreated for 1 h with or without 50 μg/mL of cell permeable inhibitors for caspase-3 (Z-DEVD-fmk), caspase-8 (Z-IETD-fmk), or a general caspase inhibitor Z-VAD-fmk, followed by treatments with 20 ng/mL TRAIL for an additional 4 h. Shown are western blots for the indicated proteins. (D) DR4 cleav- age is not inhibited by lysosome or proteasome inhibi- tors. Cells were pretreated for 1 h with Z-VAD-fmk (50 μg/mL), Bortezomib (20 nM) or chloroquine (50 μM), followed by treatments with 20 ng/mL TRAIL for an additional 4 h. DR4 and DR5 proteins were detected as in (C). Caspase activity is indicated by the cleav- age of PARP. (E) Purified recombinant DR4 and DR5 protein were incubated for the indicated times at 37°C with recombinant active caspase-3 or caspase-8 at a final concentration of 5 ng/ μl in a caspase reaction buffer containing 50 mM HEPES, pH 7.5, 100 mM NaCl, 1 mM EDTA, 10% sucrose and 10 mM DTT. The resulting mixtures were analyzed by immunoblots using anti-DR4 or anti-DR5 antibodies.

Article Snippet: Recombinant human TRAIL (rhTRAIL) expressed and purified as homotrimeric protein from E. coli, and monoclonal antibodies specific to TRAIL death receptor-4 (AF347) and -5 (MAB6312) and their phycoerythrin (PE)-conjugated forms were purchased from R&D Systems (Minneapolis, MN).

Techniques: Control, Activation Assay, Western Blot, Activity Assay, Purification, Recombinant, Incubation, Concentration Assay

Journal: BMC Cancer

Article Title: The regulation of combined treatment-induced cell death with recombinant TRAIL and bortezomib through TRAIL signaling in TRAIL-resistant cells

doi: 10.1186/s12885-018-4352-3

Figure Lengend Snippet: ILz:rhTRAIL showed the highest cell death inducing ability in TRAIL susceptible cells. Indicated cells were cultured onto a 96-well plate and treated with varying recombinant TRAIL proteins: rmT, recombinant mouse TRAIL from Peprotech (rmTRAIL); rhT, recombinant human TRAIL from Peprotech (rhTRAIL); ILz:T, isoleucine zipper hexamerization motif containing recombinant human TRAIL. ( a ) HeLa, CT26, and B16F10 cells were treated with varying recombinant TRAIL proteins (100 ng/ml). After 16 h, cell survival was examined by XTT assay. ( b ) Three human cell lines (Jurkat, MDA-MB-231, and HEK 293) and two murine cell lines (BMK and 4 T1) were treated with varying recombinant TRAIL proteins (100 ng/ml). To investigate TRAIL-susceptibility, cell death was analyzed by XTT assay 24 h after treatment. The relative values of the XTT assay were examined after comparing the results to untreated controls. * p < 0.05, compared with untreated controls by Student’s t -test

Article Snippet: Recombinant human and mouse TRAIL proteins (rhTRAIL and rmTRAIL) were purchased from PeproTech (PeproTech Korea, Seoul, South Korea): rhTRAIL (310–04); rmTRAIL (315–19).

Techniques: Cell Culture, Recombinant, XTT Assay

Journal: BMC Cancer

Article Title: The regulation of combined treatment-induced cell death with recombinant TRAIL and bortezomib through TRAIL signaling in TRAIL-resistant cells

doi: 10.1186/s12885-018-4352-3

Figure Lengend Snippet: ILz:rhTRAIL showed the highest cell death inducing ability in the combination treatment of recombinant TRAIL and bortezomib. Cells were cultured onto 96-well plates with 80% to 90% of confluency. Cell death rates were analyzed by XTT assay 24 h after the indicated treatment. The relative values of the XTT assay were examined following comparison to untreated controls. ( a ) Each cell was treated with 100 ng/ml of recombinant TRAIL protein and a distinctive amount of bortezomib as indicated: rmT, recombinant mouse TRAIL from Peprotech (rmTRAIL); rhT, recombinant human TRAIL from Peprotech (rhTRAIL); ILz:T, isoleucine zipper hexamerization motif containing recombinant human TRAIL (ILz:rhTRAIL). ( b ) Two TRAIL-resistant cell lines (B16F10 and CT26) were treated with fixed amounts of bortezomib (50 and 100 nM) and serially increasing amounts of ILz:rhTRAIL. ( c ) Two TRAIL-sensitive cell lines (HEK 293 and MDA-MB-231) were treated with fixed amounts of bortezomib (25 and 50 nM) and serially increasing amounts of ILz:rhTRAIL: bort, bortezomib. * and ** p < 0.05 and p < 0.1 by Student’s t -test, respectively

Article Snippet: Recombinant human and mouse TRAIL proteins (rhTRAIL and rmTRAIL) were purchased from PeproTech (PeproTech Korea, Seoul, South Korea): rhTRAIL (310–04); rmTRAIL (315–19).

Techniques: Recombinant, Cell Culture, XTT Assay, Comparison

Journal: BMC Cancer

Article Title: The regulation of combined treatment-induced cell death with recombinant TRAIL and bortezomib through TRAIL signaling in TRAIL-resistant cells

doi: 10.1186/s12885-018-4352-3

Figure Lengend Snippet: Combined treatment-induced cell death was inhibited by anti-TRAIL antibody and pan-caspase inhibitor. ( a ) After the pre-treatment of anti-TRAIL antibody (1 μg/ml) for 1 h, B16F10 and CT26 cells were treated with the indicated amount of bortezomib (50 nM) and varying amounts of ILz:rhTRAIL: ILz:T, ILz:rhTRAIL; bort, bortezomib; anti-T, anti-TRAIL antibody. Cell death was analyzed by XTT assay 24 h after treatment. ( b ) B16F10 and CT26 cells were cultured onto a 96-well plate and treated with ILz:rhTRAIL (100 ng/ml) and bortezomib (100 nM) with or without z-VAD-fmk (50 μM) pre-treatment for 1 h. After 24 h, cell death was assayed by XTT: ILz:T, ILz:rhTRAIL; bort, bortezomib; z-VAD, z-VAD-fmk. * p < 0.05 by Student’s t -test. ( c ) B16F10 cells were stained with propidium iodide (PI) and Annexin V using FITC Annexin V Apoptosis Detection Kit (BD Biosciences). Cells were harvested after trypsin treatment and the stained populations were analyzed by Flow cytometry (FACSCalibur™, BD Biosciences, US) using BD CellQuest™ program. The populations (%) are marked in the figures

Article Snippet: Recombinant human and mouse TRAIL proteins (rhTRAIL and rmTRAIL) were purchased from PeproTech (PeproTech Korea, Seoul, South Korea): rhTRAIL (310–04); rmTRAIL (315–19).

Techniques: XTT Assay, Cell Culture, Staining, Flow Cytometry