The expression levels of HMGB1, alpha smooth muscle actin (alpha-SMA), osteopontin (OPN), smooth muscle myosin heavy chain (SM-MHC), embryonic smooth muscle myosin heavy chain (Smemb), TXA2, PAR-1 and AT1 receptor were evaluated by Western blot analyses. Iba1-positive cells and apoptotic ...
Next, we investigated whether the cytokine-induced destabilization of the vascular structures could be prevented by inhibiting both TNFα and TGFß signaling (Fig.3). TNFα signaling was inhibited by adding a neutralizing monoclonal antibody against human TNFα. TGFß was inhibited with the small...
The sections were incubated with anti-HMGB1 mouse monoclonal antibody (R&D Systems, Minneapolis, MN, USA), anti-alpha smooth muscle actin (α-SMA) rabbit polyclonal antibody (Abcam, Cambridge, UK) or anti-Iba1 rabbit polyclonal antibody (Wako, Osaka, Japan). The sections were also counter...
with a monoclonal antibody raised against SM α-actin for 1 h in the dark at 4 °C prior to mounting in Vectashield and examination by confocal microscopy (Micro Radiance, Bio- Rad, UK). Staining of cells for von Willebrand factor failed ...
SIK1 activity was determined by Western blot as the degree of phosphorylation of its T182 residue (p182) using a specific phospho-antibody. (B) Na+,K+-ATPase activity was determined in OK cells transiently transfected with SIK1 wild type (WT), K56M mutant (which renders the kinase ...
Scheme of muscle actomyosin ATPase reaction including structural model. + indicates the plus end of actin. Boxes with solid lines and broken lines represent the strong- and weak-affinity states, respectively. The powerstroke is considered to occur mainly in the transition from the A–M–ADP–Pi...
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(Fig.6b). Fibronectin and α-smooth muscle actin (SMA) were observed in vesicular structures with three layers of mutually exclusive expression profiles: an inner layer of fibronectin+endothelial cells and a middle layer of vesicular smooth muscle cells (α-SMA+) surrounded by an outer layer of...
In injured hearts, CFs are activated by pro-fibrotic signalling such as transforming growth factor-b (TGF-b) and Rho-associated kinase (ROCK) pathways, leading to high-level expression of smooth muscle a-actin (aSMA) and extra- cellular matrix (ECM) proteins, and culminating in pathological ...
Briefly, the cytoplasmic extracted from cells were used as an antigen source in a sandwich ELISA with a primary antihistone mouse monoclonal antibody coated to the microtiter plate and a second anti-DNA mouse monoclonal antibody coupled to peroxidase. The amount of peroxidase retained in the immuno...