Journal: Journal of neuroscience methods

Article Title: Microelectrode clusters enable therapeutic deep brain stimulation without noticeable side-effects in a rodent model of Parkinson's disease.

doi: 10.1016/j.jneumeth.2021.109399

Figure Lengend Snippet: Fig. 8. Immunofluorescent staining of tissue reactions around the microelectrode cluster array. (A) Visualization of NeuN (neurons; red), ED1 (activated microglia; green) and DAPI (all nuclei; blue) staining in the implantation area. Dashed line encircles the approximate electrode cluster area. Rectangular box delineates the area enlarged in B. (B) High magnification image showing NeuN and ED1 staining near a single microelectrode in an area with high neuronal density (DAPI was omitted from this panel for enhanced clarity of the microglial and neuronal density). (C) GFAP (astrocytes) and (D) RECA (blood vessel)-staining of the same area in a section 80 µm below the section shown in A and B. All scale bars are 50 µm.

Article Snippet: The following antibodies were used: i) rabbit anti-neuronal nuclei (NeuN expressed in neuronal nuclei, 1:500, #Rb 104225, Abcam, USA), ii) mouse anti-RECA (rat endothelial cell antigen expressed by endothelial cells lining blood vessel, 1:1000, #MCA970R, Bio-Rad/ Serotec), iii) mouse anti-CD68/ED1 (marker for activated microglial cells, 1∶250, #MCA341R,Bio-Rad/ Serotec, UK), and iv) rabbit anti-glial fibrillary acid protein (GFAP, astrocytic cytoskeletal protein; 1∶5000, #Z0334, Dako, Denmark).

Techniques: Staining

Journal: Scientific Reports

Article Title: Tranilast-induced stress alleviation in solid tumors improves the efficacy of chemo- and nanotherapeutics in a size-independent manner

doi: 10.1038/srep46140

Figure Lengend Snippet: Representative images from histological analysis ( A ). Endothelial cell-specific marker CD31 (red), biotinylated tomato lectin (green) and co-expression (yellow) indicating the presence of perfused vessels, and ( B ). Expression of CD31 (red) indicating blood vessels in 4T1 and MCF10CA1a tumors. Quantification of blood vessel diameter ( C ), fraction of perfused vessels ( D ), and CD31 ( E ) area fractions in control and tranilast-treated tumors. Vessel diameter was significantly increased in both tumor models ( p = 0.038, 4T1 and p = 0.04, MCF10CA1a, n = 6–8), resulting in a significant increase in the perfused vessel fraction ( p = 0.027, 4T1 and p < 0.001, MCF10CA1a, n = 6–8), whereas CD31 area fraction was unaffected by tranilast in both tumor models. Asterisks indicate a statistically significant difference between compared groups. Scale bar: 100 μm. ( F ). Quantification of doxorubicin concentration in control versus tranilast-treated MCF10CA1a breast tumors, liver, heart and spleen tissues from corresponding CD1 nude mice. Doxorubicin (225 μg) was administered by intravenously (i.v.) via tail vein injection to the animals 4 hours prior to sacrifice. Doxorubicin concentration in tissue sample homogenates was determined by quantification of fluorescence intensity (Ex.: 470 nm, Em: 590 nm). Data represent the average + S.E. values and (*) indicates statistically significant differences between compared groups (n = 7, p < 0.05). Tranilast improved the delivery of doxorubicin in tumors by 2.5 times (p = 0.005).

Article Snippet: For blood vessel perfusion analysis, mice were slowly injected with 100 μl of 1 mg/ml biotinylated lycopersicon esculentum lectin (Vector Labs) via intracardiac injection 7 minutes prior to euthanization and tumor removal.

Techniques: Marker, Expressing, Concentration Assay, Injection, Fluorescence

Journal: Contrast Media & Molecular Imaging

Article Title: Sensitivity of Multiphase Pseudocontinuous Arterial Spin Labelling (MP pCASL) Magnetic Resonance Imaging for Measuring Brain and Tumour Blood Flow in Mice

doi: 10.1155/2018/4580919

Figure Lengend Snippet: Application of the MP pCASL sequence in mice with intracerebral U87 glioma at the day 28 time point. (a) T 1 -weighted image showing no evidence of gadolinium enhancement in the tumour injected (left) hemisphere. (b) Immunohistochemical image showing an overlay of tumour area (red) combined from tissue sections spanning 500 µ m corresponding to the MRI slice in (a). (c) CBF maps showing reduction in local CBF (arrowheads) (d-e). Immunohistochemical image showing CD31 staining of vessels (brown) in tumour-injected striatum (d) and contralateral striatum (e). (f) Graph showing CBF values in the tumour-bearing (black) and contralateral (white) striatum. CBF is reduced in the tumour-bearing compared with the contralateral striatum at day 28 time point ( p =0.02). Scale bars = 20 µ m.

Article Snippet: Sections were stained for the blood vessel marker CD31 (AF3628, R&D systems, Abingdon, UK) according to the method described by Andreou et al. [ ].

Techniques: Sequencing, Injection, Immunohistochemical staining, Staining

Journal: Nature

Article Title: Endothelial–cell FAK targeting sensitizes tumours to DNA–damaging therapy

doi: 10.1038/nature13541

Figure Lengend Snippet: a–d, Double immunostaining of B16F0 (a, b) and CMT19T (c, d) tumour sections from mice treated or not with doxorubicin (Dox.) or irradiation (Irrad.) for the apoptotic marker cleaved caspase 3 (CC3; green; a, c) or the proliferation marker Ki67 (green; b, d), and the endothelial marker PECAM (red). DAPI (blue) provides a nuclear marker. Bar charts show quantitation at 48 h post-treatment cessation of the mean number of blood vessels that are within CC3-positive tumour cell niches + s.e.m. (a, c; n = 3 mice per group) or the percentage of Ki67-positive perivascular tumour cells + s.e.m. (b, d; n = 5 mice per group). e, Conditioned media from untreated (−) and doxorubicin-treated (+) endothelial cells were applied to B16F0 cell cultures and tumour-cell survival was measured. n = 9 technical replicates. f, Conditioned medium from non-irradiated (−) or irradiated (+) endothelial cells was applied to irradiated CMT19T cells and tumour-cell survival was measured in MTS assays at 4 and 5 days. Bar charts show mean tumour-cell survival + s.e.m. according to corrected absorbance readings of MTS assays. n = 12 technical replicates. A490 nm, absorbance at 490 nm. WT, wild type. Arrowheads indicate CC3-positive perivascular tumour cells; arrows indicate Ki67-positive perivascular tumour cells. Scale bars, 100 μm (a, c); 50 mm (b, d). †P = 0.1, *P < 0.05, ***P < 0.001, Student's t-test.

Article Snippet: Immunostaining Sections of human lymphoma or mouse tumours were immunostained for endothelial cells using either anti-PECAM antibody (MEC13.3; BD Biosciences, 553370) or rat anti-endomucin (V.7C7;SantaCruz, SC-65495) in combination with either rabbit anti-FAK antibody (Cell Signaling, 3285), anti-cleaved caspase 3 (Cell Signaling, 9661), rabbit anti-Ki67 (Abcam, AB15580), or rabbit anti-p65 subunit of NF-κB (D14E12; Cell Signaling, 8242S).

Techniques: Double Immunostaining, Irradiation, Marker, Quantitation Assay