smoothelin  (Novus Biologicals)

Journal: Science Advances

Article Title: Regional differences in three-dimensional fiber organization, smooth muscle cell phenotype, and contractility in the pregnant mouse cervix

doi: 10.1126/sciadv.adr3530

Figure Lengend Snippet: ( A ) To visualize SMCs, transverse sections of whole cervices from days 12, 15, 17, and 19 gravid mice ( n ≥ 3) were immunofluoresently stained with specific markers for a contractile (MYH11 and SMTN) or synthetic phenotype (RBP1 and MYH10). Sections were also stained with CNN1 and α-SMA, which are markers of contractile smooth muscle and myofibroblasts. The grayscale bar (left) is a schematic representation of markers associated with a particular SMC phenotype. Green indicates positive immunostaining for each marker; blue indicates nuclei counterstained with 4′,6-diamidino-2-phenylindole (DAPI). ( B ) Control immunoglobulin G (IgG; left) was used as a control for the primary antibodies, and primary antibodies were omitted as controls for the secondary antibodies. Representative dual immunostaining of MYH10 (green) and MYH11 (red). Scale bars, 1000 μm; inset scale bars, 60 μm. Inset depicts high-power view of select regions (dashed box) of interest, which show longitudinal- and circumferentially oriented contractile SMCs, as well as synthetic SMCs in the broad subepithelial region and diffusely throughout the stroma.

Article Snippet: Sections were incubated overnight at 4°C with blocking solution containing the following primary antibodies: calponin (1:200 dilution; Abcam, ab46794), MYH10 (1:50; Cell Signaling Technologies, 3404), MYH11 (1:125; Abcam, ab224804), RBP1 (1:100, Abcam ab154881), α-SMA (1:200 dilution; Cell Signaling Technologies, 19245), or smoothelin (1:100; Novus Biologicals, NBP2-37931).

Techniques: Staining, Immunostaining, Marker, Control