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puma  (Novus Biologicals)
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Novus Biologicals puma
Figure 3. Ectopic overexpression of Survivin fails to suppress nutlin-3a cytotoxicity in wild-type glioma cells. A, Immunoblots were performed 48 hours after transfection and 72 hours after treatment either with DMSO or nutlin-3a. Restoration of Survivin did not <t>prevent</t> <t>p53,</t> MDM2, p21, <t>PUMA</t> and cleaved Caspase 3 induction after treatment with nutlin-3a; B, Induction of apoptosis in DMSO (CT) and nutlin-3a (N) treated cells 48 hours after transfection either with empty pcDNA3 (empty) or plasmid expression vector pcDNA3-Survivin (survivin) and 72 h after treatment. Apoptosis was measured by surface Annexin V staining and flow cytometry as described in ‘‘Patients, materials and methods’’. Average of a total of three independent assays 6 sd. empty: empty vector; Survivin: Survivin-expressing vector. doi:10.1371/journal.pone.0018588.g003
Puma, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 3. Ectopic overexpression of Survivin fails to suppress nutlin-3a cytotoxicity in wild-type glioma cells. A, Immunoblots were performed 48 hours after transfection and 72 hours after treatment either with DMSO or nutlin-3a. Restoration of Survivin did not prevent p53, MDM2, p21, PUMA and cleaved Caspase 3 induction after treatment with nutlin-3a; B, Induction of apoptosis in DMSO (CT) and nutlin-3a (N) treated cells 48 hours after transfection either with empty pcDNA3 (empty) or plasmid expression vector pcDNA3-Survivin (survivin) and 72 h after treatment. Apoptosis was measured by surface Annexin V staining and flow cytometry as described in ‘‘Patients, materials and methods’’. Average of a total of three independent assays 6 sd. empty: empty vector; Survivin: Survivin-expressing vector. doi:10.1371/journal.pone.0018588.g003

Journal: PloS one

Article Title: Activation of p53 by nutlin-3a induces apoptosis and cellular senescence in human glioblastoma multiforme.

doi: 10.1371/journal.pone.0018588

Figure Lengend Snippet: Figure 3. Ectopic overexpression of Survivin fails to suppress nutlin-3a cytotoxicity in wild-type glioma cells. A, Immunoblots were performed 48 hours after transfection and 72 hours after treatment either with DMSO or nutlin-3a. Restoration of Survivin did not prevent p53, MDM2, p21, PUMA and cleaved Caspase 3 induction after treatment with nutlin-3a; B, Induction of apoptosis in DMSO (CT) and nutlin-3a (N) treated cells 48 hours after transfection either with empty pcDNA3 (empty) or plasmid expression vector pcDNA3-Survivin (survivin) and 72 h after treatment. Apoptosis was measured by surface Annexin V staining and flow cytometry as described in ‘‘Patients, materials and methods’’. Average of a total of three independent assays 6 sd. empty: empty vector; Survivin: Survivin-expressing vector. doi:10.1371/journal.pone.0018588.g003

Article Snippet: Primary antibodies used were p53 (Neomarkers, Fremont, CA), p21 (Santa Cruz Biotechnology, Santa Cruz, CA), MDM2, PUMA and Noxa (Abcam, Cambridge, UK), Survivin (Novus Biologicals, Littleton, CO), and S6, pS6 and cleaved Caspase 3 (Cell Signaling Technology, Beverly, MA).

Techniques: Over Expression, Western Blot, Transfection, Plasmid Preparation, Expressing, Staining, Flow Cytometry