Journal: Frontiers in Cellular Neuroscience

Article Title: Crystallin Alpha-B Overexpression as a Possible Marker of Reactive Astrogliosis in Human Cerebral Contusions

doi: 10.3389/fncel.2022.838551

Figure Lengend Snippet: Glial fibrillary acidic protein (GFAP) and crystallin alpha-B (CRYAB) expression in the cortical gray matter of human brain tissue after traumatic brain injury (TBI). (A–D) GFAP expression in gray matter. (E–H) CRYAB expression in gray matter. (A,B,E,F) Control tissue. (C,D,G,H) TBI tissue. (A,C,E,G) Microphotographs with objective lens 10×/0.25, scale bar: 200 μm. (B,D,F,H) Enlargement of the box shown with dashed lines, with objective lens 40×/0.75, scale bar: 50 μm. The white arrow indicates a cell with astrocyte morphology. The black arrow indicates a cell with pyramidal neuron morphology.

Article Snippet: The adherence to non-specific antigens was blocked with normal goat serum for CRYAB and normal horse serum for NeuN and GFAP (Vectastain Elite ABC-HRP kits: products No. PK-6101 rabbit and PK-6102 mouse, respectively; Vector Labs), for 40 min. Tissue was incubated overnight in 1:100 diluted primary antibody Anti-phospho-α-B Crystallin Serine 59 produced in rabbit (Product No. C7990, Sigma-Aldrich, Merck).

Techniques: Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Crystallin Alpha-B Overexpression as a Possible Marker of Reactive Astrogliosis in Human Cerebral Contusions

doi: 10.3389/fncel.2022.838551

Figure Lengend Snippet: GFAP and CRYAB expression in the subcortical white matter of human brain tissue after TBI. (A–D) GFAP expression in white matter. (E–H) CRYAB expression in white matter. (A,B,E,F) Control tissue. (C,D,G,H) TBI tissue. (A,C,E,G) Microphotographs with objective lens 10×/0.25, scale bar: 200 μm. (B,D,F,H) Enlargement of the box shown with dashed lines, with objective lens 40×/0,75, scale bar: 50 μm.

Article Snippet: The adherence to non-specific antigens was blocked with normal goat serum for CRYAB and normal horse serum for NeuN and GFAP (Vectastain Elite ABC-HRP kits: products No. PK-6101 rabbit and PK-6102 mouse, respectively; Vector Labs), for 40 min. Tissue was incubated overnight in 1:100 diluted primary antibody Anti-phospho-α-B Crystallin Serine 59 produced in rabbit (Product No. C7990, Sigma-Aldrich, Merck).

Techniques: Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Crystallin Alpha-B Overexpression as a Possible Marker of Reactive Astrogliosis in Human Cerebral Contusions

doi: 10.3389/fncel.2022.838551

Figure Lengend Snippet: Detail of the astrocytic labeling of GFAP and CRYAB in contused brain tissue. (A,B) GFAP expression in the white matter of contused brain tissue. (A) Objective lens 40×/0.75. (B) Enlargement of the box shown with dashed lines, with objective lens 100×/1.3. (C,D) CRYAB expression in the white matter of contused brain tissue. (C) Objective lens 40×/0.75. (D) Enlargement of the box shown with dashed lines, with objective lens 100×/1.3.

Article Snippet: The adherence to non-specific antigens was blocked with normal goat serum for CRYAB and normal horse serum for NeuN and GFAP (Vectastain Elite ABC-HRP kits: products No. PK-6101 rabbit and PK-6102 mouse, respectively; Vector Labs), for 40 min. Tissue was incubated overnight in 1:100 diluted primary antibody Anti-phospho-α-B Crystallin Serine 59 produced in rabbit (Product No. C7990, Sigma-Aldrich, Merck).

Techniques: Labeling, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Crystallin Alpha-B Overexpression as a Possible Marker of Reactive Astrogliosis in Human Cerebral Contusions

doi: 10.3389/fncel.2022.838551

Figure Lengend Snippet: Immunofluorescent colocalization of GFAP and CRYAB expression in astrocytes of the gray-white matter transition of the cerebral cortex after human TBI. Microphotographs of CRYAB/GFAP/MERGE double labeling with 40×/1.3 objective lens and additional 1.0× zoom, scale bar: 100 μm. A CRYAB-positive cell with pyramidal morphology, located approximately in layer V of the contused cerebral cortex, is shown in the lower right box. Microphotographs with 40×/1.3 objective lens and additional 1.5× zoom, scale bar: 20 μm.

Article Snippet: The adherence to non-specific antigens was blocked with normal goat serum for CRYAB and normal horse serum for NeuN and GFAP (Vectastain Elite ABC-HRP kits: products No. PK-6101 rabbit and PK-6102 mouse, respectively; Vector Labs), for 40 min. Tissue was incubated overnight in 1:100 diluted primary antibody Anti-phospho-α-B Crystallin Serine 59 produced in rabbit (Product No. C7990, Sigma-Aldrich, Merck).

Techniques: Expressing, Labeling

Journal: Frontiers in Cellular Neuroscience

Article Title: Crystallin Alpha-B Overexpression as a Possible Marker of Reactive Astrogliosis in Human Cerebral Contusions

doi: 10.3389/fncel.2022.838551

Figure Lengend Snippet: Quantitative analysis of GFAP- and CRYAB-positive cells in contused brain tissue using DAB-immunohistochemistry. (A) Scatterplot of CRYAB-positive cells in human postmortem and contused brain tissue. (B) Scatterplot of GFAP-positive cells in human postmortem and contused brain tissue. (C) Comparison of the number of GFAP- and CRYAB-positive astrocytes in human contused brain tissue. Cortical sectors: SUPRA (between layers II and III), INFRA (specifically in layer V), TRANS (transition between layer VI and subcortical white matter), and DWM (deep white matter). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The adherence to non-specific antigens was blocked with normal goat serum for CRYAB and normal horse serum for NeuN and GFAP (Vectastain Elite ABC-HRP kits: products No. PK-6101 rabbit and PK-6102 mouse, respectively; Vector Labs), for 40 min. Tissue was incubated overnight in 1:100 diluted primary antibody Anti-phospho-α-B Crystallin Serine 59 produced in rabbit (Product No. C7990, Sigma-Aldrich, Merck).

Techniques: Immunohistochemistry

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Merkel Cell Carcinoma Expresses Vasculogenic Mimicry: Demonstration in Patients and Experimental Manipulation in Xenografts

doi: 10.1038/labinvest.2014.99

Figure Lengend Snippet: Biomarkers employed for identification of VM versus true angiogenesis.

Article Snippet: All sections were incubated overnight with primary antibodies at room temperature (~25°C) followed by 2-hour incubation with HRP-conjugated goat anti-rabbit (for Ulex, human and mouse CD31, CD144 and laminin), horse anti-goat (for VEGFR-1) and horse anti-mouse (for Nodal) secondary antibodies (all Vector Laboratories, Burlingame, CA) at room temperature.

Techniques:

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Merkel Cell Carcinoma Expresses Vasculogenic Mimicry: Demonstration in Patients and Experimental Manipulation in Xenografts

doi: 10.1038/labinvest.2014.99

Figure Lengend Snippet: WaGa and MKL-1-derived xenograft tumors contained CD31 − anastomosing networks that were enhanced with PAS staining, and were associated with staining for laminin (Lam), CD144, Nodal and VEGFR-1. Note the density and complexity of laminin + structures in comparison to CD31 + vessels (CD31 and Lam panels original magnification, 200×; PAS, CD144, Nodal, and VEGFR-1 original magnification, 1000×).

Article Snippet: All sections were incubated overnight with primary antibodies at room temperature (~25°C) followed by 2-hour incubation with HRP-conjugated goat anti-rabbit (for Ulex, human and mouse CD31, CD144 and laminin), horse anti-goat (for VEGFR-1) and horse anti-mouse (for Nodal) secondary antibodies (all Vector Laboratories, Burlingame, CA) at room temperature.

Techniques: Derivative Assay, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Merkel Cell Carcinoma Expresses Vasculogenic Mimicry: Demonstration in Patients and Experimental Manipulation in Xenografts

doi: 10.1038/labinvest.2014.99

Figure Lengend Snippet: MKL-1 tumors treated with vehicle, carboplatin and etoposide stained for CD31 (upper panels; original magnification, 100×) and laminin (Lam) (middle panels; original magnification, 100×); rectangular regions are representative fields enlarged for clarity (original magnification, 1000×). Note the marked and widespread increase in laminin + networks after chemotherapy. A similar picture is observed in WaGa derived tumors, although less diffusely (data not shown). A statistically significant increase in laminin immunoreactivity after carboplatin is observed in both cell lines, and after etoposide in MKL-1 derived tumors (graphs, lower panels).

Article Snippet: All sections were incubated overnight with primary antibodies at room temperature (~25°C) followed by 2-hour incubation with HRP-conjugated goat anti-rabbit (for Ulex, human and mouse CD31, CD144 and laminin), horse anti-goat (for VEGFR-1) and horse anti-mouse (for Nodal) secondary antibodies (all Vector Laboratories, Burlingame, CA) at room temperature.

Techniques: Staining, Derivative Assay

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Merkel Cell Carcinoma Expresses Vasculogenic Mimicry: Demonstration in Patients and Experimental Manipulation in Xenografts

doi: 10.1038/labinvest.2014.99

Figure Lengend Snippet: Relative LAMA3 (left), LAMB3 (center, left), LAMC2 (center, right) and CD144 (right) mRNA expression (mean ± s.e.m.) by carboplatin- and etoposide-resistant versus vehicle-treated MKL-1 (top) and WaGa (bottom) cells, as determined by real-time quantitative RT-PCR. Established human umbilical vein endothelial cells (HUVEC) served as a positive control. Data are representative of n=3 independent experiments. P<0.05, ** P<0.01, *** P<0.001).

Article Snippet: All sections were incubated overnight with primary antibodies at room temperature (~25°C) followed by 2-hour incubation with HRP-conjugated goat anti-rabbit (for Ulex, human and mouse CD31, CD144 and laminin), horse anti-goat (for VEGFR-1) and horse anti-mouse (for Nodal) secondary antibodies (all Vector Laboratories, Burlingame, CA) at room temperature.

Techniques: Expressing, Quantitative RT-PCR, Positive Control

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Merkel Cell Carcinoma Expresses Vasculogenic Mimicry: Demonstration in Patients and Experimental Manipulation in Xenografts

doi: 10.1038/labinvest.2014.99

Figure Lengend Snippet: PAS-positive networks also are demonstrable by laminin IHC, and a minority of these are associated with CD31 staining in adjacent sections original magnification, 100×; inset, 1000×). Double IF labeling for CD31 and CD144 demonstrates an architectural pattern of CD144 positivity similar to that seen with laminin, and distinct from the comparatively few and discrete CD31 + vessels (original magnification, 200×)

Article Snippet: All sections were incubated overnight with primary antibodies at room temperature (~25°C) followed by 2-hour incubation with HRP-conjugated goat anti-rabbit (for Ulex, human and mouse CD31, CD144 and laminin), horse anti-goat (for VEGFR-1) and horse anti-mouse (for Nodal) secondary antibodies (all Vector Laboratories, Burlingame, CA) at room temperature.

Techniques: Staining, Labeling