Figure 1
Figure 1. XN potentiates the TNF-induced apoptosis in chronic myeloid leukemia cells. (A) XN potentiates the TNF-induced apoptosis in chronic myeloid leukemia cell line KBM-5 as determined by live/dead assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 24 hours. The cells were stained with a live/dead assay reagent for 30 minutes and then analyzed under a fluorescence microscope. (B) XN increases TNF-induced apoptosis in human multiple myeloma U-266 cells as determined by live/dead assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 24 hours. The cells were stained with a live/dead assay reagent for 30 minutes and then analyzed under a fluorescence microscope. (C) XN potentiates TNF-induced apoptosis in KBM-5 cells as determined by annexin V assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 6 hours. The cells were incubated with a fluorescein isothiocyanate-conjugated annexin V antibody and then analyzed using flow cytometry. (D) XN potentiates TNF-induced apoptosis in KBM-5 cells as determined using a TUNEL assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 12 hours. The cells were stained for TUNEL-positive cells and then analyzed using flow cytometry. (E) XN potentiates TNF-induced apoptosis in KBM-5 cells as determined by caspase-3 activation. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for the indicated times. Whole-cell extracts were prepared and analyzed using Western blotting with an anti-PARP antibody. (F) XN suppresses TNF-induced tumor-cell invasion. H1299 cells (2.5 × 104 cells/mL) were seeded in the top chamber of a Matrigel invasion chamber system overnight in the absence of serum and then treated with 50 μM XN. After incubation, the cells were treated with TNF in the presence of 1% serum and then assayed for invasion. The results are expressed as the fold activity of the untreated control.

XN potentiates the TNF-induced apoptosis in chronic myeloid leukemia cells. (A) XN potentiates the TNF-induced apoptosis in chronic myeloid leukemia cell line KBM-5 as determined by live/dead assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 24 hours. The cells were stained with a live/dead assay reagent for 30 minutes and then analyzed under a fluorescence microscope. (B) XN increases TNF-induced apoptosis in human multiple myeloma U-266 cells as determined by live/dead assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 24 hours. The cells were stained with a live/dead assay reagent for 30 minutes and then analyzed under a fluorescence microscope. (C) XN potentiates TNF-induced apoptosis in KBM-5 cells as determined by annexin V assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 6 hours. The cells were incubated with a fluorescein isothiocyanate-conjugated annexin V antibody and then analyzed using flow cytometry. (D) XN potentiates TNF-induced apoptosis in KBM-5 cells as determined using a TUNEL assay. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for 12 hours. The cells were stained for TUNEL-positive cells and then analyzed using flow cytometry. (E) XN potentiates TNF-induced apoptosis in KBM-5 cells as determined by caspase-3 activation. Cells (106/mL) were pretreated with 50 μM XN for 4 hours and then incubated with 1 nM TNF for the indicated times. Whole-cell extracts were prepared and analyzed using Western blotting with an anti-PARP antibody. (F) XN suppresses TNF-induced tumor-cell invasion. H1299 cells (2.5 × 104 cells/mL) were seeded in the top chamber of a Matrigel invasion chamber system overnight in the absence of serum and then treated with 50 μM XN. After incubation, the cells were treated with TNF in the presence of 1% serum and then assayed for invasion. The results are expressed as the fold activity of the untreated control.

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