Journal: Oncology letters

Article Title: SMOC1 silencing suppresses the angiotensin II-induced myocardial fibrosis of mouse myocardial fibroblasts via affecting the BMP2/Smad pathway.

doi: 10.3892/ol.2018.8989

Figure Lengend Snippet: Figure 4. SMOC1 silencing mitigates the oxidative stress in MFBs induced by Ang II. ELISA was performed to evaluate the (A) MDA content, (B) LDH content and (C) SOD activity in MFBs, MFBs transfected with an empty vector, and Ang II‑induced MFBs transfected with an empty vector or si‑SMOC1. *P<0.05 and **P<0.01 vs. NC; ^P<0.05 and ^^P<0.01 vs. Ang II+NC. SMOC1, SPARC‑related modular calcium binding 1; MFBs, myocardial fibroblasts; Ang II, angiotensin II; MDA, malondialdehyde; LDH, lactate dehydrogenase; SOD, superoxide dismutase; si, small interfering RNA; NC, negative control.

Article Snippet: The expression of TGF‐β1, COL-I and COL-III were determined by ELISA kits: Mouse TGF‐β1 ELISA Kit (E‐EL‐M0051c; Elabscience, Wuhan, Hubei, China), COL1 ELISA kit (E‐EL‐M0325c; Elabscience) and COL‐III ELISA kit (E‐EL‐M0316; Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, Transfection, Plasmid Preparation, Binding Assay, Small Interfering RNA, Negative Control

Journal: Oncology letters

Article Title: SMOC1 silencing suppresses the angiotensin II-induced myocardial fibrosis of mouse myocardial fibroblasts via affecting the BMP2/Smad pathway.

doi: 10.3892/ol.2018.8989

Figure Lengend Snippet: Figure 5. SMOC1 silencing downregulates the expression levels of fibrosis‑associated proteins. (A) ELISA was performed to measure the TGF‑β1, COL‑I and COL‑III expression in MFBs, MFBs transfected with an empty vector, and Ang II‑induced MFBs transfected with an empty vector or si‑SMOC1. (B) Reverse transcription‑quantitative polymerase chain reaction and (C) western blot analysis were performed on the expression levels of FN, TGF‑β1, COL‑I and COL‑III in MFBs, MFBs transfected with an empty vector, and Ang II‑induced MFBs transfected with an empty vector or si‑SMOC1. *P<0.05, **P<0.01, and ***P<0.001 vs. NC; ^P<0.05 and ^^P<0.01 vs. Ang II+NC. SMOC1, SPARC‑related modular calcium binding 1; FN, fibronectin; TGF‑β1, transforming growth factor β1; COL, collagen; Ang II, angiotensin II; MFBs, myocardial fibroblasts; si, small interfering RNA; NC, negative control.

Article Snippet: The expression of TGF‐β1, COL-I and COL-III were determined by ELISA kits: Mouse TGF‐β1 ELISA Kit (E‐EL‐M0051c; Elabscience, Wuhan, Hubei, China), COL1 ELISA kit (E‐EL‐M0325c; Elabscience) and COL‐III ELISA kit (E‐EL‐M0316; Elabscience).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Polymerase Chain Reaction, Western Blot, Binding Assay, Small Interfering RNA, Negative Control

Journal: Applied Materials Today

Article Title: 3D bioprinting of multilayered scaffolds with spatially differentiated ADMSCs for rotator cuff tendon-to-bone interface regeneration

doi: 10.1016/j.apmt.2022.101510

Figure Lengend Snippet: Fig. 6. IHC staining of the deposition of collagen types I, II, and III at the tendon-to-bone interfaces at 6 weeks (A) and 12 weeks (B) after the infraspinatus tendon-to- bone interface reconstruction surgeries (scale bar: 100 µm; T: tendon; B: bone).

Article Snippet: The sections were then incubated overnight at 4 ◦C with anti-collagen type I primary antibody (Col I, NB600–408, Novus Biologicals), anti-collagen type II primary antibody (Col II, NB600–844, Novus Biologicals), and anti-collagen type III primary antibody (Col III, NBP1–05,119, Novus Biologicals).

Techniques: Immunohistochemistry

Journal: bioRxiv

Article Title: Effect of therapeutic ultrasound on the mechanical and biological properties of fibroblasts

doi: 10.1101/2021.11.22.469508

Figure Lengend Snippet: Protein concentration in cell culture supernatants of ligament fibroblasts. Values were measured on the 6 th day and 10 th day after stimulation period for type I collagen, type III collagen, and fibronectin.

Article Snippet: We used rat collagen type I (E-EL-R0233), rat collagen type III (E-EL-R0235), and rat fibronectin (E-EL-R0578) Elabscience® ELISA kits.

Techniques: Protein Concentration, Cell Culture

Journal: Molecular Therapy. Nucleic Acids

Article Title: USP10 Targeted Self-Deliverable siRNA to Prevent Scarring in the Cornea

doi: 10.1016/j.omtn.2020.07.032

Figure Lengend Snippet: Immunohistochemical Analysis of Collagen III after Wounding (A–C) Frozen sections of corneas 6 weeks after wounding were immunostained for collagen III (green), DAPI (blue). (A) UnWnd, (B) Wnd-NTC with magnified inset, and (C) Wnd-US09 with magnified inset. Scale bar, 0.5 mm. (D) Compared to UnWnd, Wnd-NTC demonstrated a 276.2-fold increase in collagen III immunostaining (p < 0.01), which was reduced by 71.7% (p < 0.05) with US09 treatment. The comparison between UnWnd and Wnd-US09 was not significant. (E) By qPCR, days 1 and 2 combined, compared to UnWnd, Wnd-NTC demonstrated a 4.26-fold increase in USP10 gene expression (p < 0.001). Compared to Wnd-NTC, USP10 expression with Wnd-US09 treatment was reduced by 56.8% (p < 0.01). N = 4 rabbits per condition. At 6 weeks, compared to UnWnd, Wnd-NTC demonstrated a 35.7-fold increase in USP10 gene expression (p < 0.05). Compared to Wnd-NTC, USP10 expression with Wnd-US09 treatment was reduced by 91.2% (p < 0.05). N = 6 rabbits per condition.

Article Snippet: On the next day sections were rehydrated in PBS for 15 min, treated with blocking buffer (10% normal goat serum in PBS, Jackson ImmunoResearch Laboratories) for 20 min, and then incubated with primary antibodies (FN-EDA [Sigma, F6140], collagen III [Novus Biologicals, NBP105119B], α-SMA [Sigma, C6198], CD45 Thermo Fisher Scientific, MA5-28392]) at 1:250 for 1 h in a moist chamber at room temperature (RT).

Techniques: Immunohistochemical staining, Immunostaining, Comparison, Gene Expression, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: USP10 Targeted Self-Deliverable siRNA to Prevent Scarring in the Cornea

doi: 10.1016/j.omtn.2020.07.032

Figure Lengend Snippet: Immunohistochemical Analysis of α-SMA, Cell Proliferation, and Thickness after Wounding (A–C) Frozen sections of corneas 6 weeks after wounding were immunostained for α-SMA (green), DAPI (blue). (A) UnWnd, (B) Wnd-NTC with magnified inset, and (C) Wnd-US09 with magnified inset. Scale bar, 0.5 mm. (D) Compared to UnWnd, Wnd-NTC demonstrated a 5.77-fold increase in α-SMA immunostaining (p < 0.05). Compared to Wnd-NTC, α-SMA immunostaining after Wnd-US09 treatment was reduced by 83.6% (p < 0.05). The comparison between UnWnd and Wnd-US09 was not significant. (E and F) Cell proliferation was analyzed by the object counter plugin in ImageJ software. “Inside the wound” is denoted by the anterior cornea demarcated by the collagen III scar. “Outside the wound” is the posterior cornea beneath the scar. These counts were normalized by the total area of each portion to generate a nuclei density measurement. (E) Compared to UnWnd, Wnd-NTC demonstrated a 1.61-fold increase in cell proliferation (p < 0.01). Compared to Wnd-NTC, cell proliferation after Wnd-US09 treatment was reduced by 29.9% (p < 0.05). The comparison between UnWnd and Wnd-US09 was not significant. (F) Cell proliferation below the scar, in the stroma down to the endothelium. All relationships were not significant. (G) Corneal thickness was measured at pixel resolution in these thresholded images as the distance across the nonzero region, and thickness is averaged across the entire cornea. Wnd-NTC trended toward a slight decrease in thickness (p = 0.05). Wnd-US09 treatment restored corneal thickness to non-wounded parameters. N = 6 rabbits per condition.

Article Snippet: On the next day sections were rehydrated in PBS for 15 min, treated with blocking buffer (10% normal goat serum in PBS, Jackson ImmunoResearch Laboratories) for 20 min, and then incubated with primary antibodies (FN-EDA [Sigma, F6140], collagen III [Novus Biologicals, NBP105119B], α-SMA [Sigma, C6198], CD45 Thermo Fisher Scientific, MA5-28392]) at 1:250 for 1 h in a moist chamber at room temperature (RT).

Techniques: Immunohistochemical staining, Immunostaining, Comparison, Software