Journal: International journal of molecular medicine

Article Title: Ischemic preconditioning increases GSK-3β/β-catenin levels and ameliorates liver ischemia/reperfusion injury in rats.

doi: 10.3892/ijmm.2015.2153

Figure Lengend Snippet: Figure 7. (A) mRNA and (B) protein expression levels of Bcl-2 and survivin. *P<0.01 vs. sham-operated group; #P<0.05 vs. ischemia/reperfusion (I/R) group, values are the mean ± SD; n=10 per group. IPC, ischemic preconditioning.

Article Snippet: The proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with appropriate gel concentrations (10% for β-catenin, GSK-3β, p-GSK-3β and VEGF; 15% for Bcl-2) and then electroblotted onto polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA, USA) for 2 h. The membranes were then incubated overnight at 4 ̊C with antibodies raised against β-actin (sc-47778; 1:1,000), GSK-3β (sc-9166; 1:800), p-GSK-3β (sc-11757; 1:600), β-catenin (sc-7963; 1:500), VEGF (sc-7269; 1:600), Bcl-2 (sc-7382; 1:600) and survivin (sc-17779; 1:500; all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

Techniques: Expressing

Journal: Molecular Cancer Therapeutics

Article Title: Targeting FGFR/PDGFR/VEGFR Impairs Tumor Growth, Angiogenesis, and Metastasis by Effects on Tumor Cells, Endothelial Cells, and Pericytes in Pancreatic Cancer

doi: 10.1158/1535-7163.mct-11-0312

Figure Lengend Snippet: Figure 1. Effects of TKI258 on pancreatic cancer cells in vitro. A, structure of the FGFR/PDGFR/VEGFR inhibitor TKI258 (Reprinted with friendly permission from Novartis Oncology). B, Western blot analysis of signaling intermediates. Incubation of pancreatic cancer cells with TKI258 for 2 and 20 hours led to inhibition of constitutive and FGF-1–induced ERK phosphorylation. C, MTT assay of MiaPaCa2 showed improved growth inhibition upon combination of gemcitabine and TKI258 after 48 hours (#, P < 0.05) and 72 hours (*, P < 0.05). Bars, SEM. D, migration of pancreatic cancer cells upon stimulation with FGF-1 was significantly induced (#, P < 0.05). Treatment with TKI258 impaired constitutive and FGF-1–induced motility after 24 hours (*, P < 0.05). Bars, SEM. E, inhibition of FGFR/PDGFR/VEGFR with TKI258 for 24 hours diminished expression of N-cadherin and survivin in a dose-dependent manner. Results are shown for MiaPaCa2; similar results were obtained from BxPC-3 and HPAF-II (except for Fig. 1C). OD, optical density.

Article Snippet: Membranes were sequentially probed to indicated signaling intermediates with antibodies against phosphoAktSer473, Akt, phospho-ERKThr202/Tyr204, ERK, phosphoSTAT3Tyr705, STAT3, N-Cadherin, E-Cadherin, survivin, phospho-c-RafSer289/296/301, phospho-c-RafSer338, c-Raf, phospho-FAKTyr925, FAK, FGFR3, EGF-R, andHsp90 (Cell Signaling Technologies), and phospho-VEGFR2, VEGFR2, PDGFRa, PDGFRb, and b-actin (Santa Cruz Biotechnologies).

Techniques: In Vitro, Western Blot, Incubation, Inhibition, Phospho-proteomics, MTT Assay, Migration, Expressing

Journal: Molecular Cancer Therapeutics

Article Title: Targeting FGFR/PDGFR/VEGFR Impairs Tumor Growth, Angiogenesis, and Metastasis by Effects on Tumor Cells, Endothelial Cells, and Pericytes in Pancreatic Cancer

doi: 10.1158/1535-7163.mct-11-0312

Figure Lengend Snippet: Figure 5. Impact of TKI258 on growth of established tumors in vivo. A, L3.6pl human pancreatic cancer cells were subcutaneously injected into the right flank of nude mice (n ¼ 5 per group). Treatment (TKI258, 30 mg/kg/d) was started when tumors reached a size of 400 mm3. After 4 days of treatment, tumors did not differ in terms of tumor volume, but after 6 days, a significant growth inhibition with TKI258 therapy was observed (*, P < 0.05). Bars, SEM. B, in the orthotopic model, treatment of established tumor was initiated on day 15 after tumor cell injection (n ¼ 8 per group). Treatment with TKI258 (30 mg/kg/d) led to a significant increase in survival rate of mice as shown in the Kaplan–Meier analyses (P < 0.01 vs. control). C, Western blot analyses of tumors from the orthotopic model show a decrease in expression of VEGFR2, PDGFRb, FGFR3, EGFR, survivin, and Hsp90 upon therapy with TKI258. In contrast, expression of PDGFRa was increased compared with controls. D, treatment with TKI258 led to a significant increase in Tie2 expression in orthotopic tumors (#, P < 0.05). Bars, SEM. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Membranes were sequentially probed to indicated signaling intermediates with antibodies against phosphoAktSer473, Akt, phospho-ERKThr202/Tyr204, ERK, phosphoSTAT3Tyr705, STAT3, N-Cadherin, E-Cadherin, survivin, phospho-c-RafSer289/296/301, phospho-c-RafSer338, c-Raf, phospho-FAKTyr925, FAK, FGFR3, EGF-R, andHsp90 (Cell Signaling Technologies), and phospho-VEGFR2, VEGFR2, PDGFRa, PDGFRb, and b-actin (Santa Cruz Biotechnologies).

Techniques: In Vivo, Injection, Inhibition, Control, Western Blot, Expressing

Journal: BMC Cancer

Article Title: Characterization of STAT3 activation and expression in canine and human osteosarcoma

doi: 10.1186/1471-2407-9-81

Figure Lengend Snippet: STAT3 siRNA induces downregulation of STAT3 and its downstream targets with subsequent loss of viability and apoptosis of canine OSA cells . A) Canine OSA cell lines OSA8 and OSA16 were transfected with Lipofectamine 2000 alone, 50 pMol scrambled siRNA, or 50 pMol STAT3 siRNA and collected 48 hours post transfection. Protein lysates were generated and separated via SDS-PAGE. Western blotting for STAT3, VEGF, survivin and β-actin was performed. B) OSA8 and OSA16 were transfected with Lipofectamine 2000, scrambled siRNA, or STAT3 siRNA at 0 and 48 hours. Cell viability was assessed at 0, 72, or 96 hours post transfection using the Wst-1 assay. Values are reported as percentage of control wells. C) OSA8 and OSA16 were transfected with Lipofectamine 2000, scrambled siRNA, or STAT3 siRNA and generation of active caspase-3/7 was assessed 48 hours post transfection using the SensoLyte ® Homogeneous AMC Caspase-3/7 Assay kit. D) OSA8 cells were left untreated or transfected with STAT3 siRNA or scrambled siRNA and evaluated by digital photography 72 hours post transfection. *p < 0.05

Article Snippet: The membranes were then incubated overnight with anti-p-STAT3 (Y705, Cell Signaling Technology, Danvers, MA), anti-p-Src (Y418, Invitrogen, Carlsbad, CA), anti-PARP (BD Biosciences, San Jose, CA), anti-VEGF (Calbiochem, Gibbstown, NJ), or anti-survivin antibody (Novus Biologicals, Littleton, CO).

Techniques: Transfection, Generated, SDS Page, Western Blot, WST-1 Assay, Control

Journal: BMC Cancer

Article Title: Characterization of STAT3 activation and expression in canine and human osteosarcoma

doi: 10.1186/1471-2407-9-81

Figure Lengend Snippet: Inhibition of Src or STAT3 leads to PARP cleavage and downregulates survivin expression in OSA cell lines . A) Canine or B) human OSA cell lines were treated for 24 or 48 hours with DMSO, SU6656, or LLL3. Cells were collected and protein lysates were separated via SDS-PAGE. Western blotting for survivin, PARP and β-actin was performed. The anti-PARP antibody utilized in our laboratory has been applied to experiments with canine cell lines and recognizes a 113 kDa intact PARP protein and a 23 kDa cleaved PARP fragment .

Article Snippet: The membranes were then incubated overnight with anti-p-STAT3 (Y705, Cell Signaling Technology, Danvers, MA), anti-p-Src (Y418, Invitrogen, Carlsbad, CA), anti-PARP (BD Biosciences, San Jose, CA), anti-VEGF (Calbiochem, Gibbstown, NJ), or anti-survivin antibody (Novus Biologicals, Littleton, CO).

Techniques: Inhibition, Expressing, SDS Page, Western Blot

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: Immunohistochemical staining of Survivin and fluorescent TUNEL staining in human gliomas ( A–G ). A primary glioblastoma showed a high level (++++) expression of nuclear Survivin ( A ). High staining score (++++) of cytoplasmic Survivin was detected in another primary glioblastoma ( B ). Moderate expression level (++) of cytoplasmic Survivin was detected in a pre-existing low-graded (grade II) glioma ( C ), which was obviously lower than that in its paired secondary glioblastoma with a staining score of ++++ ( D ). High-level (++++) expression of nuclear Survivin was detected in a secondary glioblastoma ( E ) and its matched pre-existing grade II glioma ( F ). Three apoptotic cells with clear nuclear staining (green color) were examined in a primary glioblastoma ( G ).

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Immunohistochemical staining, Staining, TUNEL Assay, Expressing

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: Status of Survivin expression and apoptotic index in primary GBMs

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: Status of Survivin expression and apoptotic index in secondary GBMs

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: The expression of cytoplasmic and nuclear Survivin in 15 paired secondary GBMs and pre-existing lesions a

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Expressing

Journal: BioMed Research International

Article Title: Improvement of Inflammation through Antioxidant Pathway of Gardeniae Fructus 50% EtOH Extract (GE) from Acute Reflux Esophagitis Rats

doi: 10.1155/2020/4826176

Figure Lengend Snippet: Western blot analysis of (a) survivin, (b) cytochrome c, (c) caspase-3, (d) Bax, and (e) Bcl-2 expressions. Nor: normal rats, Con: reflux esophagitis control rats, GL: GE 50 mg/kg treated reflux esophagitis rats, and GH: GE 100 mg/kg treated reflux esophagitis rats. (f) All data are expressed means ± SEM, ( n = 6) rats per group. Significance: ∗ p < 0.05, ∗∗ p < 0.01 versus RE control rat values.

Article Snippet: Mouse monoclonal antibodies against survivin (1 : 1,000, NB 500-205) were purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Western Blot, Reflux, Control