Journal: Signal Transduction and Targeted Therapy

Article Title: The β-catenin/YAP signaling axis is a key regulator of melanoma-associated fibroblasts

doi: 10.1038/s41392-019-0100-7

Figure Lengend Snippet: SK-MEL-24; GFP/Fb spheroid and SK-MEL-24; bcat-GFP/Fb spheroid cultures were grown for 96 h. Representative bright-field images of SK-MEL-24; GFP/Fb spheroid a – d and SK-MEL-24; bcat-GFP/Fb spheroid cultures e – h were taken using a Carl Zeiss Axiovert 100 TV inverted microscope at ×5 magnification at 24, 48, 72, and 96 h. i Ki67 immunofluorescence staining (red) of SK-MEL-24; GFP/Fb and SK-MEL-24; bcat-GFP/Fb spheroids embedded in paraffin. The nuclei were stained with DAPI (blue). Scale bar: 200 μm. j Quantification of Ki67-positive cells per square millimeter of SK-MEL-24; GFP/Fb spheroid sections and SK-MEL-24; bcat-GFP/Fb spheroid sections. A minimum of ten randomly selected fields was counted for each spheroid type. k Flow cytometry was used to analyze apoptosis and death of SK-MEL-24 melanoma cells in both SK-MEL-24; GFP/Fb spheroids and SK-MEL-24; bcat-GFP/Fb spheroids using annexin V and PI staining. l Percentages of SK-MEL-24 cell subpopulations based on PI and annexin V staining in SK-MEL-24; GFP/Fb spheroids and SK-MEL-24; bcat-GFP/Fb spheroids are shown. m – p α-SMA m–n and FSP-1 o – p immunostaining was performed on 5-μm-thick paraffin sections of SK-MEL-24; GFP/Fb and SK-MEL-24; bcat-GFP/Fb spheroids. Pictures were taken using a Nikon microscope at ×20 magnification. Scale bar: 200 μm. q Collagen content in SK-MEL-24; GFP/Fb spheroids and SK-MEL-24; bcat-GFP/Fb spheroids was visualized by picrosirius red staining. Pictures were taken using an inverted light microscope at ×20 magnification. r Quantification of collagen content was performed by collagen extraction and colorimetric measurement. Collagen content was normalized to total protein content for each sample. s – v Fibronectin s – t and vimentin u – v immunostaining was performed on 5-μm-thick paraffin sections of SK-MEL-24; GFP/Fb and SK-MEL-24; bcat-GFP/Fb spheroids. Pictures were taken using a Nikon microscope at ×20 magnification. Scale bar: 200 μm. w – z Photographs were taken at ×40 magnification. Scale bar: 100 μm.

Article Snippet: Picrosirius red staining was performed according to the manufacturer’s instructions (American MasterTech, Lodi, CA).

Techniques: Inverted Microscopy, Immunofluorescence, Staining, Flow Cytometry, Immunostaining, Microscopy, Light Microscopy, Extraction

Journal: Reproductive Sciences

Article Title: Cervix Stromal Cells and the Progesterone Receptor A Isoform Mediate Effects of Progesterone for Prepartum Remodeling

doi: 10.1177/1933719118820462

Figure Lengend Snippet: Top: Photomicrographs of progesterone receptor (PR) stained cells and counterstained cell nuclei (CN) in cervix sections from nonpregnant (NP), pregnant (days 15 and 18 postbreeding), and day of birth postpartum (PP) mice. Scale bar is 25 µm. Middle: Density of PR cells normalized to CN per area to account for variability in cell nuclei density due to heterogeneity of tissue morphology within and among sections in individuals, as well as within and among groups. Data are the mean ± SE (n = 5-21/group). Bottom: As inversely related to cross-linked collagen in the extracellular matrix,2 optical density of birefringence of picrosirius red-stained cervix sections cells was normalized to CN per area. Data are mean ± SE (n = 4-10/group; *P < .05 vs NP mice or **vs NP, day 15 and day 16 postbreeding groups by 1-way ANOVA). See Methods for details about staining and analyses. SE indicates standard error; ANOVA, analysis of variance.

Article Snippet: Additional sections were stained with a picrosirius red stain kit (PK-4000; Polysciences, Warrington, Pennsylvania) to assess extracellular collagen organization and counterstained with hematoxylin (SH26; Fisher Scientific, Asheville, NC) as described previously.

Techniques: Staining

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA miR-29b regulates diabetic aortic remodeling and stiffening

doi: 10.1016/j.omtn.2021.02.021

Figure Lengend Snippet: Diabetic aortae exhibit medial fibrosis, increased elastin fragmentation, Mmp2 expression, and activity (A) Representative images of aortic cross sections from 20-week-old mice stained for collagen via picrosirius red (red = collagen) and anti-collagen I immunofluorescence antibody (red = collagen), elastin (autofluorescence of green elastic lamella; arrows exemplarily indicate thinning and breaks), Mmp2 (red), and elastolytic activity ( in situ zymography). Original magnifications are 400×. Scale bar represents 50 μm. (B) Total collagen content per aortic dry weight. n = 5/group. Values are mean ± SEM. ∗p < 0.05 versus +/db control. (C) Elastin fragmentation index quantified from 3 high-power fields of 5 different aortas per group. Values are mean ± SEM. ∗p < 0.05 versus +/db control (D) Representative images of human aortic sections stained for collagen via anti-collagen I immunofluorescence antibody, and for elastin (via autofluorescence of the elastic lamella; arrows exemplarily indicate thinning and breaks). Original magnifications are 400×. Scale bar represents 100 μm. (E) Elastin fragmentation index quantified from 3 high-power fields of 5 different aortas per group. Values are mean ± SEM. ∗p < 0.05 versus non-diabetic.

Article Snippet: Aortic cross sections (7 μm) were stained using the Picrosirius Red Stain kit (American MasterTech, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Activity Assay, Staining, Immunofluorescence, In Situ, Zymography, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA miR-29b regulates diabetic aortic remodeling and stiffening

doi: 10.1016/j.omtn.2021.02.021

Figure Lengend Snippet: miR-29b inhibition de-represses extracellular matrix target genes inducing aortic fibrosis, elastin fragmentation, and aortic stiffening (A) Aortic expression levels of miR-29b target genes Col1a1 and Mmp2 in anti-miR-29b treated +/db mice versus scr controls. Values are mean ± SEM and expressed as fold change relative to the mean expression level in scr controls (=1; dotted line). ∗p < 0.05 versus +/db. n = 5/group. (B) Representative images of aortic cross sections from 20-week-old mice stained for collagen via picrosirius red (red = collagen) and anti-collagen I immunofluorescence antibody (red = collagen), elastin (autofluorescence of green elastic lamella; arrows exemplarily indicate thinning and breaks), Mmp2 (red), and elastolytic activity ( in situ zymography). Original magnifications are 400×. Scale bar represents 50 μm. (C) Total collagen content per aortic dry weight. n = 4/group. Values are mean ± SEM. ∗p < 0.05 versus scr control. (D) Elastin fragmentation index quantified from 3 high-power fields of 5 different aortas per group. Values are mean ± SEM. ∗p < 0.05 versus scr control. (E) Aortic pressure diameter curves from +/db mice treated with anti-miR-29b versus scr controls. Values are mean ± SEM. ∗p < 0.05 versus scr controls. (n = 5/group). (F) PWV measurement in +/db mice treated with anti-miR-29b versus scr controls. Values are mean ± SEM. ∗p < 0.05 versus scr controls. (n = 5/group).

Article Snippet: Aortic cross sections (7 μm) were stained using the Picrosirius Red Stain kit (American MasterTech, USA) according to the manufacturer’s instructions.

Techniques: Inhibition, Expressing, Staining, Immunofluorescence, Activity Assay, In Situ, Zymography, Control

Journal: Bioengineering

Article Title: 3D-Printed Medical-Grade Polycaprolactone (mPCL) Scaffold for the Surgical Treatment of Vaginal Prolapse and Abdominal Hernias

doi: 10.3390/bioengineering10111242

Figure Lengend Snippet: Histology overview of the vaginal wall implanted with the PP mesh and mPCL mesh scaffold at three- and six-month time points. ( A – D ) H&E; ( E – H ) Masson trichrome; ( I – L ) polarised light microscopy of Picrosirius Red staining (inset images) of the new tissue formed within the PP and mPCL meshes. Red and white dashed lines: PP mesh threads and mPCL struts. White arrows indicate collagen fibers alignment around PP nesh threads. Scale bars: 100 μm.

Article Snippet: Slides were then incubated in Picrosirius Red (Picrosirius Red Stain Kit, Abcan, Melbourne, Australia, ab150681) for 60 min.

Techniques: Light Microscopy, Staining

Journal: Bioengineering

Article Title: 3D-Printed Medical-Grade Polycaprolactone (mPCL) Scaffold for the Surgical Treatment of Vaginal Prolapse and Abdominal Hernias

doi: 10.3390/bioengineering10111242

Figure Lengend Snippet: Histology overview of the abdominal wall implanted with the PP mesh and mPCL mesh scaffold at three- and six-month time points. ( A – D ) H&E; ( E – H ) Masson trichrome; ( I – L ) polarised light microscopy and Picrosirius Red staining (inset images): of the new tissue formed within the PP and mPCL meshes. Red and white dashed lines: PP mesh threads and mPCL struts. White arrows indicate collagen fibers alignment around PP nesh threads. Scale bars: 100 μm.

Article Snippet: Slides were then incubated in Picrosirius Red (Picrosirius Red Stain Kit, Abcan, Melbourne, Australia, ab150681) for 60 min.

Techniques: Light Microscopy, Staining