pbabe-puro empty vector Search Results


pbabe puro empty vector  (Addgene inc)
93
Addgene inc pbabe puro empty vector
Pbabe Puro Empty Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBabe(puro)-Omi-mCherry+(Plasmid+%2348685)/pmc03533644-49-44-47
Average 93 stars, based on 1 article reviews
pbabe puro empty vector - by Bioz Stars, 2026-08
93/100 stars
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pbabe puro empty vector control  (Addgene inc)
96
Addgene inc pbabe puro empty vector control
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Puro Empty Vector Control, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBABE-puro+(Plasmid+%231764)/pmc10870818-37-45-50
Average 96 stars, based on 1 article reviews
pbabe puro empty vector control - by Bioz Stars, 2026-08
96/100 stars
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pbabe puro  (Addgene inc)
93
Addgene inc pbabe puro
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/Twist-siRNA3+(Plasmid+%231784)/pmc11822649-193-24-34
Average 93 stars, based on 1 article reviews
pbabe puro - by Bioz Stars, 2026-08
93/100 stars
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vector pbabe puro  (Addgene inc)
92
Addgene inc vector pbabe puro
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Vector Pbabe Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBabe+K-Ras+(Plasmid+%2312544)/pmc04479446-100-13-17
Average 92 stars, based on 1 article reviews
vector pbabe puro - by Bioz Stars, 2026-08
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pbabe puro pbp empty vector  (Addgene inc)
93
Addgene inc pbabe puro pbp empty vector
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Puro Pbp Empty Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBABEpuro-gateway+(Plasmid+%2351070)/pmc04868697-156-17-21
Average 93 stars, based on 1 article reviews
pbabe puro pbp empty vector - by Bioz Stars, 2026-08
93/100 stars
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pbabe empty vector  (Addgene inc)
94
Addgene inc pbabe empty vector
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Empty Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBabe-Puro-Myr-Flag-AKT1+(Plasmid+%2315294)/pmc09856535-98-13-27
Average 94 stars, based on 1 article reviews
pbabe empty vector - by Bioz Stars, 2026-08
94/100 stars
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empty vector pbabepuro  (Addgene inc)
94
Addgene inc empty vector pbabepuro
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Empty Vector Pbabepuro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/EGFR+WT+(Plasmid+%2311011)/us09387211-154-26-31
Average 94 stars, based on 1 article reviews
empty vector pbabepuro - by Bioz Stars, 2026-08
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pbabe puro ires egfp  (Addgene inc)
93
Addgene inc pbabe puro ires egfp
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Puro Ires Egfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBabe+puro+IRES-EGFP+(Plasmid+%2314430)/pm41681946-56-20-23
Average 93 stars, based on 1 article reviews
pbabe puro ires egfp - by Bioz Stars, 2026-08
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pbabe puromycin empty vector  (Addgene inc)
93
Addgene inc pbabe puromycin empty vector
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Puromycin Empty Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBabe-Puro-BRAF-V600E+(Plasmid+%2315269)/pm22798674-67-22-40
Average 93 stars, based on 1 article reviews
pbabe puromycin empty vector - by Bioz Stars, 2026-08
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empty pbabe puro retroviral vector ev  (Addgene inc)
93
Addgene inc empty pbabe puro retroviral vector ev
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Empty Pbabe Puro Retroviral Vector Ev, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pBABEpuro-ERBB2+(Plasmid+%2340978)/pmc07338408-169-2-13
Average 93 stars, based on 1 article reviews
empty pbabe puro retroviral vector ev - by Bioz Stars, 2026-08
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pbabe puro  (OriGene)
96
OriGene pbabe puro
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Pbabe Puro, supplied by OriGene, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pbabe-puro+empty+vector/pCMV6-XL5+Mammalian+Expression+Vector/pm25619830-196-24-29
Average 96 stars, based on 1 article reviews
pbabe puro - by Bioz Stars, 2026-08
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ha hif1α p402a p564a pbabe puro  (Addgene inc)
93
Addgene inc ha hif1α p402a p564a pbabe puro
S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with <t>pBabe-Flag</t> hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or <t>pBABE-puro</t> empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).
Ha Hif1α P402a P564a Pbabe Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ha hif1α p402a p564a pbabe puro - by Bioz Stars, 2026-08
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S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with pBabe-Flag hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or pBABE-puro empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).

Journal: Cancer Research Communications

Article Title: Pim Kinase Inhibitors Increase Gilteritinib Cytotoxicity in FLT3-ITD Acute Myeloid Leukemia Through GSK-3β Activation and c-Myc and Mcl-1 Proteasomal Degradation

doi: 10.1158/2767-9764.CRC-23-0379

Figure Lengend Snippet: S159A-mutated Mcl-1 confers resistance to Mcl-1 downregulation and to apoptosis induction by gilteritinib and AZD1208 combination treatment. A, Ba/F3-ITD cells infected with pBabe-Flag hMcl-1-S159A, containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation, pBabe-Flag hMcl-1 plasmid, containing wild-type Mcl-1, or pBABE-puro empty vector were treated with either gilteritinib and AZD1208 or DMSO control, and serial samples were immunoblotted for Mcl-1 and vinculin loading control. Densitometric analysis is also shown. B, To measure Mcl-1 protein turnover, cells were pretreated with CHX for 1 hour and then treated with gilteritinib and AZD1208 (+) or DMSO control (−). Serial samples were immunoblotted for c-Myc and vinculin loading control. Densitometric analysis was performed. Mcl-1 was normalized to vinculin and 50% protein turnover timepoints were determined to be 1.75 versus more than 2 hours for gilteritinib and AZD1208 versus DMSO control for empty vector, 1.27 versus 1.6 for wild-type Mcl-1 and more than 2 hours for both for S159A Mcl-1. C, Cells infected with pBabe-Flag hMcl-1-S159A, pBabe-Flag hMcl-1 or pBABE-puro empty vector control were treated with gilteritinib and/or AZD1208, or DMSO control, for 48 hours, and apoptosis was measured. Apoptosis induction by gilteritinib and AZD1208 combination was significantly reduced in cells infected with pBABE-puroS159A compared with empty vector control (**, P < 0.001).

Article Snippet: Ba/F3-ITD cells also underwent retroviral infection with pBabe-Flag hMcl-1-S159A plasmid (Addgene plasmid # 25372; RRID:Addgene_25372 ) containing Mcl-1 with a mutation changing serine to alanine at residue 159, preventing phosphorylation , pBabe-Flag hMcl-1 plasmid (Addgene plasmid # 25371; RRID:Addgene_25371 ), containing wild-type Mcl-1 , or pBabe -puro empty vector control (Addgene plasmid # 1764; RRID:Addgene_1764 ; ref. ).

Techniques: Infection, Mutagenesis, Residue, Plasmid Preparation