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Several other applications for which the yolk is detrimental. Here, we demonstrated that Western blotting is improved significantly using the deyolking method. The evaluation of mass spectrometry-based database searches revealed that the combination of two publicly available databases yields a good identification rate. Therefore, as a fruit of the zebrafish genome project, mass spectrometrybased i
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Erulescens green fluorescent protein; FACS: Flow-assisted cell sorting; SDS: Sodium dodecyl sulfate; MTS: 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2(4-sulfophenyl)-2 H-tetrazolium; MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide; CCD: Charge-coupled device. Competing interests None of the listed authors have competing interests related to the publication of this man
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Rtant given that zebrafish as a model system in toxicology is on the rise [12]. In addition to its application in proteomics deyolking will be beneficial to other subjects such as microarray experiments targeting the gene expression pattern in the embryo proper. We have already applied the protocol successfully to such an experiment (data not shown). Therefore, the deyolking protocol will prove a
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G/ml Triphala for indicated time points were analyzed for DCF fluorescence (ROS generation) by flow cytometry after staining the cells with DCFDA. These experiments were repeated twice and obtained similar results. B) Activation of ERK, p53 and apoptosis by ROS can be abrogated by NAC. Cells were pretreated with 5 mM NAC for 1 hour and then treated with 60 g/ml Triphala for 4 h. p-ERK, p-p53 and c
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D dish to a 1.5 ml tube filled with 1 ml deyolking buffer (1/2 Ginzburg Fish Ringer [1] without Calcium: 55 mM NaCl, 1.8 mM KCl, 1.25 mM NaHCO3) by pipetting with a narrow tip so that the yolk sac is disrupted (200 tip, Sarstedt 70.760.502). Up to 100 embryos can be transferred in a 200 volume. The embryos were shaken for 5 min at 1100 rpm to dissolve the yolk (Thermomixer, Eppendorf). Cells w
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Cal significance.Paraffin sections of our patient-derived glioblastoma xenografts (15 of 22 lines) were stained for galectin-1 expression. Around half of the xenografts tested showed preferential staining at the tumor-brain interface (Figure 3). A few tumors stained in their entirety, and another subset lacked significant staining. The 2 to 4 fold change in galectin-1 mRNA expression at the tumor
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Activation of ERK with induction of apoptosis by various chemopreventive and chemotherapeutic agents [39-41]. In fact, oxidants have been shown to activate ERK by taking over the growth factor receptor signaling pathways [42-46]. Moreover, ERK may get activated in response to DNA damage and can phosphorylate p53 in vitro [23,24,47-49]. We found that exposure of Capan-2 or BxPC-3 cells with apoptos
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File1: Figure S1. Galectin-1 staining correlates with patient survival. Using a tissue microarray created at Mayo Clinic, we stained glioblastoma samples from 34 separate patients using immunohistochemistry for galectin-1. A survival analysis revealed a trend towards shorter survival in those patients harboring galectin-1 positive tumors. Abbreviations ATCC: American type culture collection; ECM: