Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways-0

D with 1 μM BMS-345541. The IKKβ activities were examined by kinase assay using GST-IκBα as a substrate. The [γ-P]-labeled IκB-α protein was visualized by autoradiography. The IKKβ activities were quantitated by ImageQuant software. The bottom panel shows a commassie blue staining of GST-IκBα to show equal amount of substrate in each reaction. BMS-345541 inhibited IKKβ activity in C8166 cells in dose-dependent manner; however, Purvalanol A had no effect on IKKβ. Kinase assay were performed as described above using 0.01, 0.1, and 1 μM of BMS-345541 and 1, 10 μM of Purvalanol A. The stained gel below is a representative of the kinase reaction.<p><b>Copyright information:</b></p><p>Taken from "Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways"</p><p>http://www.aidsrestherapy.com/content/5/1/12</p><p>AIDS Research and Therapy 2008;5():12-12.</p><p>Published online 10 Jun 2008</p><p>PMCID:PMC2483717.</p><p></p

Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways-2

Rn blot analysis using anti-IκB, phospho IκB (ser 32), p65, phospho p65 (ser 536), p50, p52, Tax and actin. Twenty five microgram of each extract was used to separate on a 4–20% SDS/PAGE. Levels of total IκB and p65 did not change between cell types, however there was a dramatic increase of phosphor-IκB and phosphor-p65 in HTLV-1 infected cells and their suppression by BMS-345541 which inhibits IKKβ activity in vivo.<p><b>Copyright information:</b></p><p>Taken from "Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways"</p><p>http://www.aidsrestherapy.com/content/5/1/12</p><p>AIDS Research and Therapy 2008;5():12-12.</p><p>Published online 10 Jun 2008</p><p>PMCID:PMC2483717.</p><p></p

Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways-4

Ned with a mixture of propidium iodide buffer followed by cell sorting analysis. The acquired FACS data were analyzed by ModFit LT software (Verity Software House, Inc.).<p><b>Copyright information:</b></p><p>Taken from "Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways"</p><p>http://www.aidsrestherapy.com/content/5/1/12</p><p>AIDS Research and Therapy 2008;5():12-12.</p><p>Published online 10 Jun 2008</p><p>PMCID:PMC2483717.</p><p></p

Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways-1

Total cell extracts were subjected to Western blot analysis for caspase-3 and PARP. β-actin Western blot was used as internal control. The results of caspase-3 were quantitated and normalized with β-actin. The ratio of c/un PARP was calculated by dividing cleaved PARP to un-cleaved PARP (data not shown). Detection of apoptosis through annexin V and PI staining. Cells were washed three times in PBS and re-suspended in binding buffer, stained with annexin V-FITC and PI for 15 minutes at room temperature. Analysis was performed on a BD FacsCalibur flow cytometer.<p><b>Copyright information:</b></p><p>Taken from "Two specific drugs, BMS-345541 and purvalanol A induce apoptosis of HTLV-1 infected cells through inhibition of the NF-kappaB and cell cycle pathways"</p><p>http://www.aidsrestherapy.com/content/5/1/12</p><p>AIDS Research and Therapy 2008;5():12-12.</p><p>Published online 10 Jun 2008</p><p>PMCID:PMC2483717.</p><p></p