Journal: Scientific Reports

Article Title: Trans -pairing between osteoclasts and osteoblasts shapes the cranial base during development

doi: 10.1038/s41598-018-38471-w

Figure Lengend Snippet: Localization of osteoclasts and osteoblasts in the basioccipital bone. ( a ) Representative micro-CT image of a mouse skull at P14 (n = 4). E, ethmoid; PS, presphenoid; BS, basisphenoid; BO, basioccipital (dorsal surface, green; parasagittal section, magenta); SOS, spheno-occipital synchondrosis; Arrows indicate the clivus. Voxel size, 40 μm. Scale bar, 5 mm. ( b ) Dorsal view of the cranial base shown in ( a ) and a scheme of the basioccipital bone. O, otic capsule; F, foramen magnum. Arrows indicate the clivus. Scale bar, 5 mm. ( c ) Safranin O staining of the cranial base in a representative section from a P3 mouse (n = 6). Arrows indicate the clivus of the basioccipital between the brainstem (pons and medulla, Po + Me) and longus capitis muscle (Mu). Pit, pituitary; Scale bar, 500 μm. ( d ) I 2 KI-contrast staining in a representative sample from a P16 mouse (n = 1). Arrows indicate the clivus. Voxel size, 10 μm. Scale bar, 500 μm. ( e ) Schematic showing the cranial base at an early postnatal stage. Asterisk, foramen magnum. BM, bone marrow. Red boxed region indicates area shown in ( f ). ( f ) Col1a1 -AcGFP and MMP-9 immunostaining in representative sections from a P2 mouse (n = 2). (Left) Fluorescence detection of AcGFP (green), MMP-9 (magenta), and DAPI staining (white) in a P2 mouse. Scale bar, 100 μm. (Right) Magnified view of the boxed area in “DAPI”. Broken lines indicate endocranial and ectocranial cortical bone layers. Arrows, osteoclasts; arrowheads, osteoblasts; BM, bone marrow between cortical bones. A, anterior; P, posterior; D, dorsal; V, ventral. Maximum intensity projection images were acquired using a 63x objective and assembled from 3 slices at 2.86 μm intervals. Scale bar, 20 μm.

Article Snippet: Sections were blocked with 1% BSA/PBS and then incubated with goat anti-mouse MMP-9 polyclonal antibody (AF909, R&D) or rabbit anti-mouse collagen type I polyclonal antibody (AB765P, Merck Millipore) overnight at 4 °C, followed by donkey anti-goat Alexa Fluor 647 or donkey anti-rabbit Alexa Fluor 488 secondary antibodies (Thermo Fisher Scientific), respectively.

Techniques: Micro-CT, Staining, Immunostaining, Fluorescence

Journal: Scientific Reports

Article Title: Trans -pairing between osteoclasts and osteoblasts shapes the cranial base during development

doi: 10.1038/s41598-018-38471-w

Figure Lengend Snippet: Mice injected with anti-RANKL antibody. Mice were injected with saline or anti-RANKL antibody at P1 and then injected with alizarin complexone at P3. The basioccipital bone was then analysed at P4 (n = 4 each group). ( a ) MMP-9 immunostaining. ( b ) TRAP activity staining. ( c ) Merge image of MMP-9 and TRAP staining. ( d ) Alizarin labelling. ( e ) Collagen type I immunostaining. ( f ) Merged image of alizarin labelling and collagen type I staining. Bone resorption (arrows) and formation (arrowheads) surfaces are indicated. Maximum intensity projection images were acquired using a 40x objective and assembled from 12 slices at 0.6 μm intervals. Scale bars, 50 μm.

Article Snippet: Sections were blocked with 1% BSA/PBS and then incubated with goat anti-mouse MMP-9 polyclonal antibody (AF909, R&D) or rabbit anti-mouse collagen type I polyclonal antibody (AB765P, Merck Millipore) overnight at 4 °C, followed by donkey anti-goat Alexa Fluor 647 or donkey anti-rabbit Alexa Fluor 488 secondary antibodies (Thermo Fisher Scientific), respectively.

Techniques: Injection, Immunostaining, Activity Assay, Staining

Journal: Biomedicines

Article Title: The Glymphatic Response to the Development of Type 2 Diabetes

doi: 10.3390/biomedicines12020401

Figure Lengend Snippet: Histological measurements: mean ± SE ( n = 18 rats per group).

Article Snippet: The following primary antibodies were used: goat anti-fibrin/fibrinogen (Accurate Chemical & Scientific, Carle Place, NY, USA; 1:1000), rabbit anti-thrombocyte (Inter-Cell Technologies, Hopewell, NJ, USA; 1:900), mouse anti-endothelial barrier antigen (EBA, Sternberger Monoclonals, Lutherville, MD, USA; 1:1000), rabbit anti-aquaporin-4 (AQP4, Abcam, Cambridge, UK; 1:1500), rabbit anti-beta Amyloid 1–42 (Aβ, Abcam, Cambridge, UK; 1:100), and mouse anti-MMP-9 (Chemicon, Temecula, CA, USA; 1:1000).

Techniques:

Journal: Biomedicines

Article Title: The Glymphatic Response to the Development of Type 2 Diabetes

doi: 10.3390/biomedicines12020401

Figure Lengend Snippet: DM-induced vascular damage. The double immunofluorescent images show intravascular fibrin (green in ( A ), arrows) and platelet (green in ( B ), arrow) deposition and peri-vascular AQP4 immunoreactivity (green in ( C ), arrow) in the hippocampus of non-DM and 3M-DM rats. Peri-vascular beta-amyloid accumulation and MMP9 immunoreactivity were detected in 3M-DM (( D ), arrows).

Article Snippet: The following primary antibodies were used: goat anti-fibrin/fibrinogen (Accurate Chemical & Scientific, Carle Place, NY, USA; 1:1000), rabbit anti-thrombocyte (Inter-Cell Technologies, Hopewell, NJ, USA; 1:900), mouse anti-endothelial barrier antigen (EBA, Sternberger Monoclonals, Lutherville, MD, USA; 1:1000), rabbit anti-aquaporin-4 (AQP4, Abcam, Cambridge, UK; 1:1500), rabbit anti-beta Amyloid 1–42 (Aβ, Abcam, Cambridge, UK; 1:100), and mouse anti-MMP-9 (Chemicon, Temecula, CA, USA; 1:1000).

Techniques:

Journal: Molecular cancer therapeutics

Article Title: NEO212 Inhibits Migration and Invasion of Glioma Stem Cells

doi: 10.1158/1535-7163.MCT-17-0591

Figure Lengend Snippet: (A) Representative images of invasion assay through a Matrigel-coated Boyden Chamber, performed with USC02 cells and 15 μM treatments. Scale bar, 200 μm. (B) After overnight treatments with 15 μM NEO212, the number of invaded cells per field was counted. The bar graph represents the average ± SEM of at least three independent experiments performed in triplicate. *, P<0.05 (relative to vehicle-treated cells). (C) After 72 h, area covered by invaded USC04 cells in the spheroid invasion assay was quantified, and represented as relative to the initial sphere. The bar graph represents the average ± SEM of three independent experiments performed in sextuplicate. *, P<0.05; **, P<0.01 (relative to vehicle-treated cells). (D) Representative images of the spheroid migration assay performed with USC04 cells at 3 μM. Scale bar, 200 μm. (E-F) Western blot analysis of matrix metalloproteinases MMP2 and MMP9 in USC02 (E) and USC04 (F) cells. The bar graphs represent the average of three independent experiments. *, P<0.05; **, P<0.01, ***, P<0.001 (relative to vehicle-treated cells); #, P< 0.05; ##, P< 0.01 (relative to TMZ-treated cells).

Article Snippet: Primary antibodies used were rabbit anti-phospho-AKT (Ser473), rabbit anti-AKT, rabbit anti-phospho-FAK (Tyr397), rabbit anti-FAK, rabbit anti-eIF2α, rabbit anti-phospho-MEK1/2 (Ser217/221), rabbit anti-MEK1/2, rabbit anti-phospho-p38-MAPK (Thr180/Tyr182), rabbit anti-p38-MAPK, rabbit anti-phospho-Src (Tyr416), rabbit anti-Src (1:1000, Cell Signaling), goat anti-phospho-eIF2α (Ser51), rabbit anti-MMP2 (1:1000, Abgent), mouse anti-MMP9 (1:500, Abgent), rabbit anti-tubulin (1:1000, BioLegend).

Techniques: Invasion Assay, Migration, Western Blot