Journal: Scientific Reports

Article Title: In vivo imaging of kidney glomeruli transplanted into the anterior chamber of the mouse eye

doi: 10.1038/srep03872

Figure Lengend Snippet: (a) and (b) Transmission electron microscopy shows interdigitating podocyte foot processes. (c) Immunogold electron microscopy reveals conserved expression of podocalyxin in podocytes from transplanted glomeruli. Control sections stained with secondary antibody only were completely negative for gold particles.

Article Snippet: Briefly, Ultrathin sections were blocked with 1% ovalbumin in PBS for one hour, followed by incubation with affinity-purified goat antibody to podocalyxin (R&D, AF1556, 1:400) and an anti-goat 10 nm gold conjugate (1:50).

Techniques: Transmission Assay, Electron Microscopy, Expressing, Staining

Journal: Scientific Reports

Article Title: Nexilin/ NEXN controls actin polymerization in smooth muscle and is regulated by myocardin family coactivators and YAP

doi: 10.1038/s41598-018-31328-2

Figure Lengend Snippet: Nexilin/ NEXN is expressed in striated and smooth muscle cells (SMCs) and correlates at the mRNA level with the smooth muscle myosin. Panel A shows RNA-Seq data (GTExPortal.org) for the top-twelve NEXN -expressing human tissues (N = 6–430, Esophagus-M: muscular layer, -GEJ: Gastroesophageal Junction). Panel B shows immunohistochemical staining for NEXN in human heart (top left), skeletal muscle (top right), esophagus (bottom left) and gall bladder (bottom right), all from the Human Protein Atlas (proteinatlas.org). Panel C shows correlation between smooth muscle myosin ( MYH11 ) and NEXN in human coronary artery (N = 133, data from the GTExPortal). Panel D shows our own double staining for Nexilin (green, ab213628) and CAV1 (red) in cross-sectioned human urinary bladder SMCs (white scale bar represents 10 µm, N = 2) imaged in a conventional fluorescence microscope.

Article Snippet: Sections were blocked with 3% BSA for 60 min at room temperature before incubation with the following primary antibodies overnight: Nexilin (1:200 in 3% BSA, Abcam, ab213628 or HPA011185, Sigma), CAV1 (1:400 in 3% BSA, Cell Signaling Technology, #3267) and YAP1 (1:200 in 3% BSA, Cell Signaling Technology, #4912).

Techniques: RNA Sequencing, Expressing, Immunohistochemical staining, Staining, Double Staining, Fluorescence, Microscopy

Journal: Scientific Reports

Article Title: Nexilin/ NEXN controls actin polymerization in smooth muscle and is regulated by myocardin family coactivators and YAP

doi: 10.1038/s41598-018-31328-2

Figure Lengend Snippet: Nexilin localizes to puncta in the cytosol and at the membrane and is excluded from caveolae domains at the membrane. Nexilin ( NEXN ) and CAV1 immunofluorescence was imaged by confocal microscopy and using two different Nexilin antibodies (ab213628 in ( A) and HPA011185 in ( B ). Both show punctate/granular distribution of Nexilin (green) in the cytoplasm and at the membrane in human bladder SMCs that overlaps poorly with CAV1 (red) staining. Correlation of the two labels (CAV1 and NEXN ) indicated poor co-localization (panel C, R = 0.073 and 0.18, for the respective NEXN antibodies). The fluorescence intensity ratio for the two labels was also plotted for three membrane profiles ( D ). Panel E shows immuno-electron microscopy for Nexilin. The gold particle on the secondary antibody appears as a black sphere with a diameter of 10 nm, and examples of dense bodies (dbo) and dense bands (dba) are shown. White arrowheads highlight gold particles.

Article Snippet: Sections were blocked with 3% BSA for 60 min at room temperature before incubation with the following primary antibodies overnight: Nexilin (1:200 in 3% BSA, Abcam, ab213628 or HPA011185, Sigma), CAV1 (1:400 in 3% BSA, Cell Signaling Technology, #3267) and YAP1 (1:200 in 3% BSA, Cell Signaling Technology, #4912).

Techniques: Membrane, Immunofluorescence, Confocal Microscopy, Staining, Fluorescence, Immuno-Electron Microscopy

Journal: Scientific Reports

Article Title: Nexilin/ NEXN controls actin polymerization in smooth muscle and is regulated by myocardin family coactivators and YAP

doi: 10.1038/s41598-018-31328-2

Figure Lengend Snippet: NEXN correlates with gene products that control and respond to changes in actin polymerization and Nexilin is reduced by depolymerization of actin. Correlations of NEXN versus all other RNAs in the top-ten NEXN expressing tissues were examined (data from GTExPortal). The sum of correlation coefficients for individual RNAs across tissues was calculated (R sum ) and the positive extreme of this distribution was plotted ( A ). Actin controlling and responding gene products represented in the extreme are highlighted in blue colors. Examples of NEXN correlations in the human coronary artery (N = 133) are shown in panels B through ( D ). P-values and Spearman Rho values are given in the respective panels. Panels E and F show mRNA data for NEXN in cultured human bladder (HBSMCs, N = 8) and coronary artery (HCASMCs, N = 9) SMCs after treatment with Latrunculin B (LatB). Panels H and I show protein data for Nexilin/ NEXN in the presence and absence of LatB (HBSMCs, 300 nM, N = 12; HCASMCs, 100 nM, N = 10). The top micrographs in panel J shows confocal imaging of YAP (red) on the left, and YAP (green) and CAV1 (red) on the right. The bottom row shows YAP (red) and Nexilin (green). The high magnification overlay at the bottom right shows partial colocalization of YAP and Nexilin at the cell membrane in yellow. All micrographs are from cross-sectioned HBSMCs and white scale bars represent 5 μm throughout.

Article Snippet: Sections were blocked with 3% BSA for 60 min at room temperature before incubation with the following primary antibodies overnight: Nexilin (1:200 in 3% BSA, Abcam, ab213628 or HPA011185, Sigma), CAV1 (1:400 in 3% BSA, Cell Signaling Technology, #3267) and YAP1 (1:200 in 3% BSA, Cell Signaling Technology, #4912).

Techniques: Control, Expressing, Cell Culture, Imaging, Membrane