Journal: The Journal of pathology

Article Title: Receptor for hyaluronan-mediated motility (RHAMM) defines an invasive niche associated with tumor progression and predicts poor outcomes in breast cancer patients.

doi: 10.1002/path.6082

Figure Lengend Snippet: Figure 4. RHAMM drives phenotypes associated with invasive breast cancer in MCF10DCIS.com (A–D) and MDA-MB-231 cell lines (E–H). MTT proliferation assay of (A) MCF10DCIS.com and (F) MDA-MB-231 parental, control, and RHAMM KO cell lines. Relative colony counts (size >50 μm) of (B) MCF10DCIS.com and (G) MDA-MB-231 parental, control, and RHAMM KO cell lines in soft agar. (C) MCF10DCIS.com parental, control, and RHAMM KO cell lines embedded in Matrigel + 20% collagen + 10 μg/ml LMW-HA and scored for invasion. Representative images of tumor spheres are shown below. (D) Representative immunoblot for vimentin in MCF10DCIS.com parental, control, and RHAMM KO cell lines. (E) Representative immunoblot for RHAMM in MDA-MB-231 Ctrl and RHAMM KO cell lines. (H) Quantification of Transwell migration assay of MDA-MB-231 control (Ctrl) and RHAMM KO cells. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001.

Article Snippet: Cells were incubated with Texas Red-labeled LMW HA (Cat #H-250R; Echelon Biosciences, Salt Lake City, UT, USA) in fresh medium for 45 min at 37 C in the dark, then washed with PBS and fixed with 2% PFA.

Techniques: Proliferation Assay, Control, Western Blot, Transwell Migration Assay

Journal: Brain Sciences

Article Title: Localization of Fibrinogen in the Vasculo-Astrocyte Interface after Cortical Contusion Injury in Mice

doi: 10.3390/brainsci7070077

Figure Lengend Snippet: Deposition of fibrinogen (Fg) in vasculo-astrocyte interface after cortical contusion injury (CCI) in mice. ( A ) Examples of astrocytes and their endfeet surrounding vessels and Fg (red) deposited in the extravascular space 14 days after sham-operation (sham) and CCI. Astrocytes are identified by expression of glial fibrillary acidic protein (GFAP, green) and vascular endothelium is defined by Lycopersicon Esculentum agglutinin tomato lectin (LEA, blue). Images of each area of interest were collected by acquiring at least 4 focal planes with increment of 0.5 µm to form a final z-stack image. The highlighted areas were enlarged 3-times (side images). Short white arrows point to Fg and astrocyte endfeet co-localization (shown in yellow). Placing the Image Pro-plus software’s line profile probes (horizontal and vertical yellow lines) on enlarged images, we obtained fluorescence intensity profiles along the X (panels are shown at the bottom of the respective enlarged images) and Y (panels are shown on the sides of the respective enlarged images) axis. Fluorescence intensity profiles show that at the area of co-localizations (shown in yellow), Fg (red) was located between the vessels (blue) and astrocyte endfeet (green). In the right horizontal panel, thin and long white arrows pointing to a vessel (blue), Fg (red), and astrocyte labeled with GFP (green) in the same vertical plane clearly show Fg between the vessel and astrocyte; ( B ) Shown are examples of z-stack images of spot co-localization after deconvolution of original images of brain samples from mice with sham-operation) or with CCI); ( C ) Average number of co-localized Fg and GFAP spots calculated after deconvolution of experimental stack images are presented for each experimental group. * p < 0.05 vs. Sham. n = 4.

Article Snippet: Texas Red-conjugated Lycopersicon Esculentum agglutinin (TXR-LEA) tomato lectin was from Vector laboratories (Burlingame, CA, USA).

Techniques: Expressing, Fluorescence, Labeling

Journal: Journal of neurochemistry

Article Title: Trafficking of green fluorescent protein tagged-vesicular acetylcholine transporter to varicosities in a cholinergic cell line.

doi: 10.1046/j.1471-4159.2001.00494.x

Figure Lengend Snippet: Fig. 4 GFP2VAChT is present in early and recycling endosomes. SN56 cells were transfected with GFP2VAChT and examined by confocal microscopy (a and c). (b) Labeling of cells in (a) with Tfn- TxR for 5 min at 378C. Arrows indicate some of the structures labeled by both GFP2VAChT and Tfn-TxR. Bar, 10 mm. (d) Label- ing of cells in (c) with Tfn-TxR for 20 min at 378C. Arrows indicate colocalization of GFP2VAChT and transferrin in recycling endo- somes. The arrowhead shows colocalization in early endosomes. Bar, 20 mm.

Article Snippet: Labeling of endosomes was also performed incubating cells with 30 mg/mL of Texas Red-labeled transferrin (Tfn-TxR; Molecular Probes) at 378C in 5% CO2 for 5 min (labeling of early endosomes) or 20 min (labeling of recycling endosomes).

Techniques: Transfection, Confocal Microscopy, Labeling