bim sirna  (Santa Cruz Biotechnology)

Journal: The Journal of Clinical Investigation

Article Title: Cotargeting DNA topoisomerase II enhances efficacy of RAS-targeted therapy in KRAS-mutant cancer models

doi: 10.1172/JCI197192

Figure Lengend Snippet: ( A ) The tested cell lines were pretreated with 10 μM MG132 for 30 minutes and then cotreated with DMSO, 500 nM sotorasib (Soto), or 250 nM adagrasib (Adag) for another 6 hours. ( B ) The indicated cell lines were treated with 500 nM sotorasib or adagrasib for 16 hours followed by the addition of 10 μg/mL CHX and then harvested at the indicated times. ( C ) The tested cell lines were pretreated with 10 μM CHIR99021 or SB216763 for 30 minutes and then cotreated with 500 nM sotorasib for an additional 16 hours. ( D and E ) Both H358 and Calu-1 cells were transfected with scrambled control, GSK3 ( D ), or FBXW7 ( E ) siRNA for 48 hours followed by treatment with 500 nM sotorasib for another 24 hours. Proteins in the treated cells were detected by Western blotting. Band intensities were quantified using ImageJ software (NIH) and plotted as a percentage of 0 time point ( B ). ( F ) Schematic illustration of KRAS G12C inhibition-induced GSK3-dependent and FBXW7-mediated degradation of Topo IIα in KRAS G12C–mutant cancer cells. G12Ci, G12C inhibitor.

Article Snippet: TOP2A siRNA (sc-36695), TOP2A shRNA plasmid (sc-36695-SH), and Bim siRNA (sc-29803) were purchased from Santa Cruz Biotechnology.

Techniques: Transfection, Control, Western Blot, Software, Inhibition, Mutagenesis

Journal: The Journal of Clinical Investigation

Article Title: Cotargeting DNA topoisomerase II enhances efficacy of RAS-targeted therapy in KRAS-mutant cancer models

doi: 10.1172/JCI197192

Figure Lengend Snippet: ( A – D ) Both H358/SR and Calu-1/SR cells expressing pLKO.1 or shTOP2A ( A and B ) or transfected with scrambled control or TOP2A siRNA for 48 hours ( C and D ) were exposed to DMSO or 10 μM sotorasib for 48 hours. The levels of Topo IIα and cleavage of PARP were detected with Western blotting ( A and C ). Annexin V–positive cells were determined with flow cytometry ( B and D ). Data are shown as the mean ± SD of triplicate determinations. ( E ) The indicated cell lines described above were exposed to DMSO or 10 μM sotorasib for 48 hours and then IF conducted to stain γ-H2AX foci. Scale bars: 25 μm, 5 μm (zoom). ( F – I ) Mice with the indicated tumors derived from H358/SR cells were treated with vehicle (V) or sotorasib (50 mg/kg, daily, og) for 24 days. Tumor sizes ( F ) and mouse body weights ( I ) were measured at the indicated time points, and at the end of the treatment, collected tumors were photographed ( G ) and weighed ( H ). Data are shown as the mean ± SEM of 6 tumors from 6 mice. Statistical differences among the treatments were determined by 1-way ANOVA.

Article Snippet: TOP2A siRNA (sc-36695), TOP2A shRNA plasmid (sc-36695-SH), and Bim siRNA (sc-29803) were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Transfection, Control, Western Blot, Flow Cytometry, Staining, Derivative Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: PLK1 inhibition enhances gemcitabine-induced apoptosis through PLK1-dependent ERK1/2-Bim and AKT1/Noxa signals in pancreatic cancer cells

doi: 10.1007/s12032-025-03062-z

Figure Lengend Snippet: Effect of targeting PLK1 by siRNA on cell viability, apoptosis and GEMCITABINE sensitivity A PANC-1 and BxPC-3 cells were exposure to series concentration of GEMCITABINE for 48 h. Cell viability was detected by MTT assay. B PLK1 siRNA transfection inhibits PLK1 expression in PANC-1 cells detected by Western blotting. C Effect of combined PLK1 siRNA and GEMCITABINE treatment on cell growth in PANC-1 cells by MTT assay. D Effect of combined PLK1 siRNA and GEMCITABINE treatment on cell apoptosis in PANC-1 cells by TUNEL assay. E Effect of combined PLK1 siRNA and GEMCITABINE treatment on PARP cleavage in PANC-1 cells. Data are means ± SEM from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: SignalSilence® PLK1 siRNA, Bim EL siRNA, Noxa siRNA, p44/42 MAPK (Erk1/2) siRNA, Akt1 siRNA, and SignalSilence® Control siRNA (CN siRNA) were purchased from Cell Signaling Technology (CST).

Techniques: Concentration Assay, MTT Assay, Transfection, Expressing, Western Blot, TUNEL Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: PLK1 inhibition enhances gemcitabine-induced apoptosis through PLK1-dependent ERK1/2-Bim and AKT1/Noxa signals in pancreatic cancer cells

doi: 10.1007/s12032-025-03062-z

Figure Lengend Snippet: Targeting PLK1 inhibits ERK1/2 and AKT1, and enhances ERK1/2-dependent Bim and AKT1-dependent Noxa expression. A PANC-1 cells were transfected with PLK1 siRNA or/and co-treated with ERK1/2 activator SDF1-α (200 ng/ml for 5 min) or SC79. ERK1/2 phosphorylation was detected by western blot assay. B PANC-1 cells were transfected with PLK1 siRNA or/and co-treated with ERK1/2 activator SDF1-α (200 ng/ml for 5 min) or SC79, the protein expression of Bim, PUMA, Noxa and AKT1 phosphorylation in PANC-1 was detected by western blot assay. C The stable PLK1 shRNA transfected PANC-1 cells were transfected with pcDNA3-PLK1 for 48 h. The protein expression of PLK1, Bim, PUMA, Noxa and ERK1/2 phosphorylation in PANC-1 was detected by western blot assay. D The stable pcDNA.3-PLK1-transfected BxPC-3 cells were transfected with AKT1 siRNA or ERK1/2 siRNA for 48 h, The protein expression of PLK1, Bim, PUMA, Noxa, AKT1 phosphorylation and ERK1/2 phosphorylation in BxPC-3 was detected by western blot assay. Data are means ± SEM from three independent experiments. * p < 0.05, ** p < 0.01,*** p < 0.001

Article Snippet: SignalSilence® PLK1 siRNA, Bim EL siRNA, Noxa siRNA, p44/42 MAPK (Erk1/2) siRNA, Akt1 siRNA, and SignalSilence® Control siRNA (CN siRNA) were purchased from Cell Signaling Technology (CST).

Techniques: Expressing, Transfection, Phospho-proteomics, Western Blot, shRNA

Journal: Medical Oncology (Northwood, London, England)

Article Title: PLK1 inhibition enhances gemcitabine-induced apoptosis through PLK1-dependent ERK1/2-Bim and AKT1/Noxa signals in pancreatic cancer cells

doi: 10.1007/s12032-025-03062-z

Figure Lengend Snippet: Targeting PLK1 induced apoptosis and enhances gemcitabine sensitivity by Bim-dependent and Noxa-dependent way in PANC-1 cells. PANC-1 cells were co-transfection with PLK1 siRNA/Bim siRNA or PLK1 siRNA/Noxa siRNA alone for 48 h, or for 16 h, then exposure to 10 μM GEM for 48 h. Cell apoptosis was detected in PANC-1 cells by TUNEL assay. * p < 0.05, ** p < 0.01

Article Snippet: SignalSilence® PLK1 siRNA, Bim EL siRNA, Noxa siRNA, p44/42 MAPK (Erk1/2) siRNA, Akt1 siRNA, and SignalSilence® Control siRNA (CN siRNA) were purchased from Cell Signaling Technology (CST).

Techniques: Cotransfection, TUNEL Assay

Journal: Medical Oncology (Northwood, London, England)

Article Title: PLK1 inhibition enhances gemcitabine-induced apoptosis through PLK1-dependent ERK1/2-Bim and AKT1/Noxa signals in pancreatic cancer cells

doi: 10.1007/s12032-025-03062-z

Figure Lengend Snippet: GEMCITABINE transiently activates ERK1/2 and PLK1 expression, and induces Bim-dependent and Noxa-dependent cell apoptosis. A PANC-1 cells were exposure to 10 μM GEMCITABINE for 5,10, 30 min, 1, 3, 6, 12 and 24 h. ERK1/2 phosphorylation, AKT1 phosphorylation and PLK1 expression was detected by western blot assay. B BxPC-3 cells were exposure to 1 μM GEMCITABINE for 5, 10, 30 min, 1, 3, 6, 12 and 24 h. ERK1/2 phosphorylation, AKT1 phosphorylation, PLK1, Bim and NOXA expression was detected by western blot assay. C PANC-1 cells were co-transfection with Bim siRNA or Noxa siRNA or Bim siRNA + Noxa siRNA for 24 h, then exposure to 10 μM GEMCITABINE for 48 h. Cell apoptosis was detected by TUNEL assay. Data are means ± SEM from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: SignalSilence® PLK1 siRNA, Bim EL siRNA, Noxa siRNA, p44/42 MAPK (Erk1/2) siRNA, Akt1 siRNA, and SignalSilence® Control siRNA (CN siRNA) were purchased from Cell Signaling Technology (CST).

Techniques: Expressing, Phospho-proteomics, Western Blot, Cotransfection, TUNEL Assay

Journal: Biomaterials Research

Article Title: Tumoricidal Efficacy of Artesunate-Eluting Microsphere: Differential Role of Bax/Bak in Orchestration of Cell Death Pathways

doi: 10.34133/bmr.0217

Figure Lengend Snippet: Bid is not required for MMC-induced cytotoxicity. HCT116 WT, Bid −/− , Bid −/− /Bid, Bid −/− /D60E, and Bid −/− /G94E cells were treated with 10, 20, or 50 μM MMC for 24 h. (A) Whole-cell extracts were analyzed with an immunoblotting assay using indicated antibodies to compare treatment effects in WT HCT116 cells and Bid −/− cells. (B) Again, whole-cell extracts were analyzed with an immunoblotting assay using indicated antibodies to compare treatment effects in WT HCT116 cells, Bid −/− cells, and Bid −/− /Bid cells, respectively. (C) To compare treatment effects in WT HCT116 cells, Bid −/− /D60E, and Bid −/− /G94E cells, whole-cell extracts were analyzed with an immunoblotting assay. (D) Before treatment with 10 μM MMC for 24 h, HC116 WT cells were transfected with Bim siRNA using Lipofectamine 3000 kit. Then, whole-cell extracts were analyzed with an immunoblotting assay using indicated antibodies.

Article Snippet: SignalSilence Bim siRNA (#6461) was used (Cell Signaling Technology, Beverly, MA).

Techniques: Western Blot, Transfection