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: Current Protocols

Article Title: Cryopreservation of Human Adult Ventricular Tissue for the Preparation of Viable Myocardial Slices

doi: 10.1002/cpz1.70068

Figure Lengend Snippet: Primary Antibodies

Article Snippet: Type III Collagen , 1:2000 , Southern Biotech , 1330‐01.

Techniques:

Journal: Current Protocols

Article Title: Cryopreservation of Human Adult Ventricular Tissue for the Preparation of Viable Myocardial Slices

doi: 10.1002/cpz1.70068

Figure Lengend Snippet: Extracellular matrix. Representative images of immunohistochemical analysis of fresh and cryopreserved cardiac tissue. Type I Collagen (Collagen‐I) is shown in orange (A) and Type III Collagen (Collagen‐III) is shown in yellow (B) . Counterstaining of nuclei was performed with DAPI (blue).

Article Snippet: Type III Collagen , 1:2000 , Southern Biotech , 1330‐01.

Techniques: Immunohistochemical 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: RSC Advances

Article Title: Thymoquinone protects against cardiac damage from doxorubicin-induced heart failure in Sprague-Dawley rats

doi: 10.1039/c8ra00975a

Figure Lengend Snippet: Fig. 4 Protein levels of TGF-b, Smad3, collagen I, collagen III, and a-SMA in cardiac tissue were determined by western blotting. Immunoblotting for TGF-b, Smad3, collagen I, collagen III, and a-SMA in cardiac tissues. Bar graph showing quantification of TGF-b, Smad3, collagen I, collagen III, and a-SMA protein expression. Data are given as the mean SEM; n ¼ 3 in each group. *p < 0.05 vs. DOX group.

Article Snippet: Primary antibodies against TGF-b (rabbit anti-TGF-b antibody, 1 : 1000; Proteintech, Wuhan, China), Smad3 (rabbit anti-Smad antibody, 1 : 1000; Proteintech), collagen I (rabbit anti-collagen I antibody, 1 : 1000; Proteintech), collagen III (rabbit anticollagen III antibody, 1 : 1000; Proteintech), a-SMA (rabbit anti-a-SMA antibody, 1 : 1000; Proteintech), P53 (rabbit antiP53 antibody, 1 : 1000; Proteintech), bcl-2(rabbit anti-bcl-2 antibody, 1 : 1000; Proteintech), anti-b-actin (1 : 1000; Cell Signaling Technology).

Techniques: Western Blot, Expressing

Journal: Journal of Thrombosis and Haemostasis

Article Title: Enhanced activity of an ADAMTS ‐13 variant (R568K/F592Y/R660K/Y661F/Y665F) against platelet agglutination in vitro and in a murine model of acute ischemic stroke

doi: 10.1111/jth.14275

Figure Lengend Snippet: Recombinant wild‐type ( WT ) ADAMTS ‐13 is only partially active in whole blood under flow. (A) Whole human blood, collected on PPACK and clexane, was perfused over a collagen‐coated surface at arterial shear rate, and the von Willebrand factor ( VWF )‐mediated capture of labeled platelets (%) was recorded over time (Control). This was repeated with the addition of either 2.5 n m ADAMTS ‐13 ( WT or gain‐of‐function [GoF]) with or without preincubation with 100 n m VWF D4‐ CK (+). As a VWF ‐negative control, blood was preincubated with an inhibitory m A b (6B4) to block the VWF –glycoprotein 1b interaction ( VWF NEG ). As an ADAMTS ‐13‐negative control, blood was preincubated with an inhibitory anti‐ ADAMTS ‐13 m A b (3H9) to inhibit endogenous ADAMTS ‐13 ( AD 13 NEG ). The curves are representative of three independent experiments. (B) For each sample, platelet coverage (%) was determined at the end (180 s) of the experiment, and pairwise t ‐tests were performed between WT and GoF ADAMTS ‐13 and their corresponding VWF D4‐ CK ‐activated samples (*** P < 0.005). Results are mean ± standard deviation ( SD ), n = 3–6. (C, D) WT ADAMTS ‐13 and the GoF variant were then tested at a range of concentrations in the same assay. For comparison, VWF ‐negative controls are included in (C). All results are given as mean ± SD, n = 3. CON , control; NS , not significant.

Article Snippet: For parallel‐flow assays, performed in the absence of coagulation, Vena8 Fluoro+ biochips (Cellix, Dublin, Ireland) were coated with 200 μg mL −1 collagen type III (Southern Biotech, Birmingham, AL, USA) and blocked with 1% bovine serum albumin (BSA) and 1 mg mL −1 glucose in HEPES buffer.

Techniques: Recombinant, Shear, Labeling, Control, Negative Control, Blocking Assay, Standard Deviation, Variant Assay, Comparison

Journal: Journal of Thrombosis and Haemostasis

Article Title: Enhanced activity of an ADAMTS ‐13 variant (R568K/F592Y/R660K/Y661F/Y665F) against platelet agglutination in vitro and in a murine model of acute ischemic stroke

doi: 10.1111/jth.14275

Figure Lengend Snippet: Gain‐of‐function (GoF) ADAMTS ‐13 partially dissolves von Willebrand factor ( VWF )/platelet‐rich pseudothrombi under flow. (A) Whole human blood, collected on citrate and containing labeled platelets, was recalcified immediately prior to perfusion over a collagen/tissue factor‐coated surface. After 9 min of continuous perfusion, large, VWF /platelet‐rich pseudothrombi (shown previously to also contain fibrin(ogen) ) were formed. At the end of this period ( t = 0 min) the channel was perfused for a further 4 min with labeled, recalcified blood, and platelet fluorescence was continuously monitored. (B, C) The assay was repeated with the addition of 5 n m ADAMTS ‐13 (wild‐type [ WT ] or GoF) during the second perfusion period. Images are representative of three independent experiments. (D) Fluorescence readings for each sample were normalized to that at t = 0 min, and are expressed as mean ± SD , n = 3. [Color figure can be viewed at http://www.wileyonlinelibrary.com ]

Article Snippet: For parallel‐flow assays, performed in the absence of coagulation, Vena8 Fluoro+ biochips (Cellix, Dublin, Ireland) were coated with 200 μg mL −1 collagen type III (Southern Biotech, Birmingham, AL, USA) and blocked with 1% bovine serum albumin (BSA) and 1 mg mL −1 glucose in HEPES buffer.

Techniques: Labeling, Fluorescence