Journal: Life Science Alliance

Article Title: Local tissue mechanics control cardiac pacemaker cell embryonic patterning

doi: 10.26508/lsa.202201799

Figure Lengend Snippet: (A) Col3A1 staining at the SAN/atria junction in representative HH18, HH30, and HH35 hearts. (B) Distribution of Col3A1, TnC, and WFA in the H35 SAN and atria. (C) Diagram of in vivo transfection strategy for mosaic calcium imaging in the intact SAN. Integrating DNA plasmids were microinjected into the pericardial space adjacent to the forming SAN at HH18 and SAN tissue was isolated for live imaging at HH30. (D) Live imaging of a memRFP/GCaMP6f-positive cell within the SAN. Data presented in were taken from cells imaged in 25 individual hearts.

Article Snippet: The following reagents were used for immunohistochemistry: MF20 (14650380; Invitrogen) (1:500); FLRT3 (PA563240; Invitrogen) (1:250); hyaluronic acid binding protein, bovine nasal cartilage, biotinylated (38591150UG; MilliporeSigma) (1:100); Wisteria floribunda lectin biotinylated (B13552; Vector Laboratories) (1:100); Col3A1 (3B2-s; DSHB) (1:250), TNC (LS-C39574-50; LSBio) (1:500); Nav1.5 (LS-C30483; LSBio) (1:500), phalloidin-647 (A22287; Invitrogen) (1:40); NCX1 (LS-C73201-100; LSBio) (1:500); streptavidin-568 (S11226; Invitrogen) (1:500); DAPI (62248; Thermo Fisher Scientific) (1:1,000); IgM (heavy chain) cross-adsorbed goat anti-mouse, Alexa Fluor 555 (A21426; Invitrogen) (1:500); IgG1 cross-adsorbed goat anti-mouse, Alexa Fluor 647 (A21240; Invitrogen) (1:500); IgG2b cross-adsorbed goat anti-mouse, Alexa Fluor 488 (A21141; Invitrogen) (1:500); IgG (H+L) cross-adsorbed goat anti-rabbit, Alexa Fluor 568 (A11011; Invitrogen) (1:500); goat anti-mouse IgG2b cross-adsorbed secondary antibody, Alexa Fluor 647 (A21242; Invitrogen) (1:500); goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody, Alexa Fluor 488 (A11008; Invitrogen) (1:500); goat anti-mouse IgM mu chain Alexa Fluor 488 (ab150121; Abcam) (1:500).

Techniques: Staining, In Vivo, Transfection, Imaging, Isolation

Journal:

Article Title: A mutant, noninhibitory plasminogen activator inhibitor type 1 decreases matrix accumulation in experimental glomerulonephritis

doi: 10.1172/JCI200318038

Figure Lengend Snippet: Immunofluorescent staining score for ECM proteins at d6. Glomerular staining for FN-EDA+ (a), laminin (b), type I collagen (c), and type III collagen (d) were lower in the PAI-1R–treated, nephritic group. *P < 0.001 compared with normal control. #P < 0.01 compared with disease control.

Article Snippet: Monoclonal mouse anti-cellular fibronectin extra domain positive (FN-EDA+) (Harlan Sera-Lab Ltd., Loughborough, United Kingdom), rabbit anti–mouse laminin (ICN Immunobiologicals, Aurora, Ohio, USA), goat anti–human type I collagen, and goat anti–human type III collagen (Southern Biotechnology Associates, Birmingham, Alabama, USA) were used as the primary Ab’s for detection of matrix components in glomeruli at d6.

Techniques: Staining

Journal: Journal of Diabetes Research

Article Title: Relationship between Urinary Liver-Type Fatty Acid-Binding Protein (L-FABP) and Sarcopenia in Spontaneously Diabetic Torii Fatty Rats

doi: 10.1155/2020/7614035

Figure Lengend Snippet: Histological evaluation of the kidney in SD and SDT fatty rats. Immunohistological staining of kidney tissues using antibodies against CD68 (a, b), α -SMA (c, d), type I collagen (e, f), and type III collagen (g, h). Histological PAS staining showing focal glomerular sclerosis (i) and semiquantitative assessment of focal glomerular sclerosis (j). Original magnification: ×100. Values are presented as the mean ± SEM. ∗ p < 0.05 vs. SD rats.

Article Snippet: The tissue specimens fixed in methyl Carnoy's solution were assessed immunohistochemically for macrophages using a mouse monoclonal antibody (ED-1) specific for CD68 (1 : 100; Abcam, Tokyo, Japan) and goat polyclonal antibodies specific for type I and III collagen (1 : 200; Southern Biotech, Birmingham, AL, USA).

Techniques: Staining

Journal: Journal of Diabetes Research

Article Title: Relationship between Urinary Liver-Type Fatty Acid-Binding Protein (L-FABP) and Sarcopenia in Spontaneously Diabetic Torii Fatty Rats

doi: 10.1155/2020/7614035

Figure Lengend Snippet: Relationships between renal morphological change and muscle strength, and weights of the soleus and extensor digitorum longus (EDL) muscles in 24-week-old SD and SDT fatty rats.

Article Snippet: The tissue specimens fixed in methyl Carnoy's solution were assessed immunohistochemically for macrophages using a mouse monoclonal antibody (ED-1) specific for CD68 (1 : 100; Abcam, Tokyo, Japan) and goat polyclonal antibodies specific for type I and III collagen (1 : 200; Southern Biotech, Birmingham, AL, USA).

Techniques: Muscles, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Secretome from In Vitro Mechanically Loaded Myoblasts Induces Tenocyte Migration, Transition to a Fibroblastic Phenotype and Suppression of Collagen Production

doi: 10.3390/ijms222313089

Figure Lengend Snippet: Col I/III production from co-cultures with and without mechanically loaded myoblasts. ( A , D ) Relative mRNA expression of Col 1a1 and Col 3a1 in tenocytes. ( B , E ) Quantification of Col 1 content in tenocyte lysate by ELISA. ( C , F ) Quantification of Col I/III content and the ratio in the cell culture medium. ( G ) Immunofluorescence staining of Col I on tenocytes. Teno ctrl: tenocytes alone, Co ctrl: tenocytes co-cultured with myoblasts, Dyn: tenocyte co-cultured with dynamically loaded myoblasts, Stat: tenocyte co-cultured with statically loaded myoblasts. Data are represented as mean ± standard deviation. n = 3. Student’s t -test was performed when comparing Teno ctrl and co ctrl. One-way ANOVA with Tukey’s multiple comparisons was performed when comparing between Co ctrl, Dyn and Stat groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Collagen I and collagen III were assessed using Rat Collagen Type I ELISA kit (Cusabio, Wuhan, China, #CSB-E08084r) and Rat Collagen Type III ELISA kit (Cusabio, Wuhan, China, #CSB-E07924r) according to the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Immunofluorescence, Staining, Standard Deviation

Journal: Translational neuroscience

Article Title: Decreased MiR-30a promotes TGF-β1-mediated arachnoid fibrosis in post-hemorrhagic hydrocephalus.

doi: 10.1515/tnsci-2020-0010

Figure Lengend Snippet: Figure 6: In the in vitro model of arachnoid cell fibrosis, the expression of miR-30a decreased, and the expression of TRAF3IP2 was upregulated. (a) Differential mRNA expression of miR-30a, TRAF3IP2, and TGF-β1 in arachnoid cells between the fibrotic group and control group (**P < 0.01 compared with the control group). (b) Differential protein expression of TRAF3IP2, TGF-β1, and α-SMA in arachnoid cells between the fibrotic and control groups (**P < 0.01 compared with the control group; *P < 0.05 compared with the control group). (c) Differential content of collagen I and collagen III in the model of arachnoid cell fibrosis (**P < 0.01 compared with the control group).

Article Snippet: ELISA kit was used to detect the TGF-β1 level (EK0514, Wuhan Boster Biological Technology, Ltd, China), collagen type I level (CSB-E08084r, CUSABIO, China), and collagen type III level (CSB-E07924r, CUSABIO, China) following the protocols constructions.

Techniques: In Vitro, Expressing, Control

Journal: Translational neuroscience

Article Title: Decreased MiR-30a promotes TGF-β1-mediated arachnoid fibrosis in post-hemorrhagic hydrocephalus.

doi: 10.1515/tnsci-2020-0010

Figure Lengend Snippet: Figure 7: MiR-30a inhibits subarachnoid fibrosis through inhibiting TRAF3IP2. (a and b) The mRNA and protein expression of miR-30a, TRAF3IP2, TGF-β1 and α-SMA from different groups (**P < 0.01 compared with the control group; ***P < 0.001 compared with the control group). (c) The content of collagen I and collagen III from different groups (***P < 0.001 compared with the control group. **P < 0.01 compared with the control group; *P < 0.05 compared with the control group).

Article Snippet: ELISA kit was used to detect the TGF-β1 level (EK0514, Wuhan Boster Biological Technology, Ltd, China), collagen type I level (CSB-E08084r, CUSABIO, China), and collagen type III level (CSB-E07924r, CUSABIO, China) following the protocols constructions.

Techniques: Expressing, Control