myh 11  (Novus Biologicals)

Journal: BioMed Research International

Article Title: The Influence of Simulated Microgravity on Purinergic Signaling Is Different between Individual Culture and Endothelial and Smooth Muscle Cell Coculture

doi: 10.1155/2014/413708

Figure Lengend Snippet: Characterization of Bovine Endothelial and Smooth Muscle Cells. The isolated cells were verified with the EC specific markers VEGFR2, VE-cadherin, PECAM-1 and the SMC specific markers calponin, SMA-α, MYH-11 by RT-PCR (a) and Western blot (b). GAPDH served as internal control. The endothelial (c) and smooth muscle cells (d) were also identified with the above-mentioned markers via fluorescent staining. The isolated endothelial cells were further examined for the typical endothelial activity of LDL up-take (e). All pictures are representative of one cow sample out of three.

Article Snippet: The membrane was blocked in TBST containing 5% BSA and incubated with anti-P2X7, P2Y1, P2Y2, P2Y11, VEGFR2, VE-cadherin, PECAM-1, calponin, SMA-α, MYH-11 (1 : 500), or GAPDH antibodies (1 : 5,000) (Santa Cruz Biotechnology, CA, USA) overnight at 4°C.

Techniques: Isolation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Activity Assay

Journal: Theranostics

Article Title: Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule

doi: 10.7150/thno.47006

Figure Lengend Snippet: Characteristics of the SMCs, EPCs, and cocultured cells. ( A ) Phase-contrast image of the bladder SMCs at P3. ( B ) The α-SMA and Myh 11 immunofluorescence staining of the SMCs were positive. ( C ) FCM showed that 98.4% of the SMCs were positive for α-SMA and Myh 11. ( D ) The functional characteristics of the EPCs were assessed by measuring their ability to uptake DiI-Ac-LDL (red fluorescence) and bind to FITC-UEA-1 (green). ( E and F ) Immunofluorescence images showing CD34 and CD31 expression in the EPCs. ( G ) Tube formation by the EPCs was assessed on Matrigel-precoated plates using PCLM. ( H ) Tube-forming EPCs were labeled with CellTracker Green (green). ( I ) The networked EPCs within the cocultured cells were stained with anti-mouse CD31 (green). ( J ) Angiogenic factor secretion of the SMCs, EPCs, and SMCs-EPCs cocultured cells was measured via enzyme-linked immunosorbent assay (n = 9). The results are expressed as means ± SD. ** p < 0.01. Nuclei were counterstained with DAPI (blue). DiI-Ac-LDL, DiI-acetylated low density lipoprotein; EPCs, endothelial progenitor cells; FCM, flow cytometry; P0, primary culture; P3, third-passage cultures; PCLM, phase-contrast light microscopy; SMCs, smooth muscle cells; UEA-I, Ulex europaeus agglutinin-1.

Article Snippet: Anti-Myh 11 (mouse monoclonal, 1:300 dilution, Novus Biologicals) and anti-α-SMA antibody (mouse monoclonal, 1:500 dilution, Abcam) were used to verify the smooth muscle origin.

Techniques: Immunofluorescence, Staining, Functional Assay, Fluorescence, Expressing, Labeling, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Light Microscopy

Journal: Theranostics

Article Title: Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule

doi: 10.7150/thno.47006

Figure Lengend Snippet: Molecular and histological analysis of reconstructed rabbit bladders. ( A ) Representative H&E and α-smooth muscle actin (α-SMA) immunohistochemical staining in retrieved bladders 1 and 3 months after cystoplasty in the 3 groups. ( B ) Induction of the smooth muscle myosin heavy chain 11 (Myh 11) expression in newly formed tissues was determined by real-time polymerase chain reaction. Values were normalized to the GAPDH levels. ※ Significant difference among the three groups at each time point after cystoplasty ( p < 0.05).

Article Snippet: Anti-Myh 11 (mouse monoclonal, 1:300 dilution, Novus Biologicals) and anti-α-SMA antibody (mouse monoclonal, 1:500 dilution, Abcam) were used to verify the smooth muscle origin.

Techniques: Immunohistochemical staining, Staining, Expressing, Real-time Polymerase Chain Reaction