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: bioRxiv

Article Title: Allele-specific expression reveals genetic drivers of tissue regeneration in mice

doi: 10.1101/2022.09.23.509223

Figure Lengend Snippet: Schematic of MRL and CAST dermal fibroblast culture from dorsal and ear skin. B. Left, fluorescent histology of cultured MRL and CAST dorsal and ear fibroblasts with immunohistochemical (IHC) staining for complement factor H (CFH) and DAPI nuclear counterstain. Right, quantification of CFH expression across in vitro conditions. C. Schematic of MRL and CAST dorsal and ear wounding for histology. D. Fluorescent histology (left) and quantification (right) of IHC staining of wounds for CFH (with DAPI nuclear counterstain). E. Schematic of wildtype mouse dorsal splinted wounding with local wound treatment with either recombinant CFH protein or phosphate-buffered saline (PBS; vehicle control) (see Methods for full details and dosing). F. Left, gross photographs of control (-CFH) and CFH-treated (+CFH) wounds; black dotted outline indicates healed wound region. Right, wound curve reflecting rate of re-epithelialization of -CFH vs. +CFH wounds over time. G. Picrosirius red connective tissue histology of -CFH and +CFH wounds and unwounded skin (UW). H. T-distributed stochastic neighbor embedding (t-SNE) plot of quantified extracellular matrix (ECM) ultrastructural parameters, based on picrosirius red histology of unwounded skin and POD 14 wounds ( G ), showing overall similarities/differences in ECM ultrastructure between conditions. Each dot represents quantified parameters from one histologic image. I. Hematoxylin and eosin (H&E) histology of POD 14 wounds and skin. Yellow dotted lines denote borders of healed wounds; white arrows indicate putative regenerating dermal appendages (hair follicles or glands) in +CFH wounds. J. Dermal thickness quantified from histology of wounds and skin. B, C, F, J. * P < 0.05 (Student’s t-test).

Article Snippet: Hematoxylin and eosin (H&E) and picrosirius red staining (using Picro Sirius Red Stain Kit, Abcam) were performed on paraffin sections, using standard protocols without modification.

Techniques: Cell Culture, Immunohistochemical staining, Immunohistochemistry, Expressing, In Vitro, Recombinant