Journal: Helminthologia

Article Title: The Synthesis of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine Kinase (GNE), α-dystroglycan, and β-galactoside α-2,3-sialyltransferase 6 (ST3Gal6) By Skeletal Muscle Cell As a Response To Infection with Trichinella Spiralis

doi: 10.2478/helm-2022-0027

Figure Lengend Snippet: Immunohistochemistry. Modified methacarn fixed sections from mouse skeletal muscles with Trichinella spiralis at days 14 and 35 post invasion (d.p.i.) were stained with rabbit polyclonal antibodies against glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine kinase (GNE), α-dystroglycan and ST3 beta-galactoside alpha-2,3-sialyltransferase 6 (ST3Gal6). Paralel sections were subjected to H&E staining to facilitate the histological orientation. Strong expressions of GNE and α-dystroglycan, and moderate expression of ST3Gal6 were observed on days 14 and 35 after invasion, suggesting these proteins as permanent characteristics of the Nurse cell of T. spiralis . The brown colour indicates positive immunohistochemical reaction, hashtag indicates the occupied sarcoplasm, star – non-occupied skeletal muscle cell, arrow – enlarged nucleus, L– larva. H&E, HRP anti-rabbit IgG, DAB. Scale bar 20 μm.

Article Snippet: The sections were then treated with a secondary antibody (ImmPress HRP anti-rabbit IgG polymer detection kit, Vector Laboratories) for 30 min, a color reaction was developed with DAB Peroxidase (HRP) Substrate Kit (Vector Laboratories) and the sections were counterstained with hematoxylin.

Techniques: Immunohistochemistry, Modification, Staining, Expressing, Immunohistochemical 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: 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

Journal: Cell reports

Article Title: Post-Golgi carriers, not lysosomes, confer lysosomal properties to pre-degradative organelles in normal and dystrophic axons

doi: 10.1016/j.celrep.2021.109034

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Subsequent staining steps were performed according to manufacturer’s instructions (Vector Laboratories), which include incubation with biotinylated secondary antibodies, complex formation with avidin and biotinylated HRP using Vectorstain ABC Kit, and reaction with Vector DAB Peroxidase Substrate.

Techniques: Plasmid Preparation, Electron Microscopy, Recombinant, Immunodetection, Fluorescence, Transfection, Expressing, Software

Journal: bioRxiv

Article Title: Chondrocyte-specific RUNX2 Overexpression Causes Chondrodysplasia During Development, but is Not Sufficient to Induce OA-like Articular Cartilage Degeneration in Adult Mice Without Injury

doi: 10.1101/470005

Figure Lengend Snippet: Chondrocyte-specific RUNX2 overexpression during development leads to chondrodysplasia. (A) Alcian blue/Alizarin red skeletal staining of E18.5 R26 Runx2/+ (Control), Col2a1 CreERT2/+ ; R26 Runx2/+ ( C2T2; R26 Runx2/+ ), and Col2a1 CreERT2/+ ; R26 Runx2/Runx2 ( C2T2; R26 Runx2/Runx2 ) embryos. (B) Femur (left), tibia (middle), and humerus (right) lengths from E18.5 Control, C2T2; R26 Runx2/+ , and C2T2; R26 Runx2/Runx2 embryos (n = 3 to 5 for all groups). * p < 0.05, ** p < 0.01, *** p < 0.001, one-way ANOVA followed by Dunnett’s multiple comparisons test. (C) Alcian blue Hematoxylin/Orange G staining of humerus growth plate sections from E18.5 Control, C2T2; R26 Runx2/+ , and C2T2; R26 Runx2/Runx2 embryos. Top panels are 5X images of the proximal humerus; middle panels are high magnification images (20X) of the proliferating and pre-hypertrophic zones (yellow boxes); bottom panels are high magnification images (20X) of the hypertrophic zone (orange boxes). (D) In situ hybridization for Col10a1 (top panels) and Mmp13 (bottom panels) on E18.5 Control, C2T2; R26 Runx2/+ , and C2T2; R26 Runx2/Runx2 proximal humerus sections. (E) TUNEL staining of proximal humerus sections from E18.5 Control, C2T2; R26 Runx2/+ , and C2T2; R26 Runx2/Runx2 embryos (5X images). (B) Quantification of TUNEL + cell number as percentage of total cell number within the growth plate of the proximal humeri of E18.5 Control, C2T2; R26 Runx2/+ , and C2T2; R26 Runx2/Runx2 embryos (n = 3 for all groups). * p < 0.05, one-way ANOVA followed by Dunnett’s multiple comparisons test.

Article Snippet: Sections were rinsed in phosphate buffered saline + 0.1% tween-20 (PBST) and incubated with ImmPress HRP anti-rabbit IgG peroxidase polymer secondary (anti-MMP13, Vector Labs) or ImmPress HRP anti-mouse IgG peroxidase polymer secondary (anti-COL10A1, Vector Labs) for 30 minutes.

Techniques: Over Expression, Staining, In Situ Hybridization, TUNEL Assay

Journal: bioRxiv

Article Title: Chondrocyte-specific RUNX2 Overexpression Causes Chondrodysplasia During Development, but is Not Sufficient to Induce OA-like Articular Cartilage Degeneration in Adult Mice Without Injury

doi: 10.1101/470005

Figure Lengend Snippet: Postnatal RUNX2 overexpression induces MMP13 expression in unmineralized articular chondrocytes prior to accelerating articular cartilage degeneration following MLI. (A) Safranin O/Fast green staining of knee joint sections from R26 Runx2/+ (Control) and Acan CreERT2/+ ; R26 Runx2/+ (RUNX2 OE) mice injected with tamoxifen at 2 months of age and subjected to sham or MLI at 2.5 months of age. Joints were harvested one month following injury. Top panels are 5X images of the knee joint; bottom panels are high magnification images (20X) of the boxed regions. (B) Cartilage degeneration in Control (n = 6) and RUNX2 OE (n = 6) mice was evaluated by modified OARSI scoring (top graph) and quantitative histomorphometric analysis (bottom graph, data normalized to contralateral sham control sample). (C) MMP13 (left panels) and COL10A1 (right panels) immunohistochemistry of knee joint sections from Control or RUNX2 OE mice subjected to sham or MLI at 2.5 months of age and harvested 1 month following injury (20X images). (D) Quantification of MMP13 + cell number as percentage of total cell number from Control and RUNX2 OE mice (MLI samples only, n = 4 for both groups). (E) Quantification of COL10A1 + cartilage area as percentage of total cartilage area from Control and RUNX2 OE mice (MLI samples only, n = 4 for both groups). * p < 0.05, Student’s t-test.

Article Snippet: Sections were rinsed in phosphate buffered saline + 0.1% tween-20 (PBST) and incubated with ImmPress HRP anti-rabbit IgG peroxidase polymer secondary (anti-MMP13, Vector Labs) or ImmPress HRP anti-mouse IgG peroxidase polymer secondary (anti-COL10A1, Vector Labs) for 30 minutes.

Techniques: Over Expression, Expressing, Staining, Injection, Modification, Immunohistochemistry