rabbit anti laminin Search Results


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  • 99
    Thermo Fisher rabbit anti laminin
    <t>Laminin</t> assembly is compromised in mammary epithelial cells lacking DG expression. ( A ) Primary mammary epithelial cells from control or ΔDG K14-Cre mice were incubated with 10 ng/ml FITC-laminin-111 overnight. DG-expressing cells (WT) showed efficient assembly of laminin, whereas DG-knockout cells (KO) showed little assembled laminin. The phase-contrast images below show the positions of the cells. ( B ) Cells derived from the immortalized DG-knockout cell line (MEpL), transfected with DG (WT) or a vector control (KO) were overlaid with 10 ng/ml FITC-laminin-111 overnight. WT cells show efficient assembly of laminin, whereas the knockout cells show no assembled laminin. The differential interference contrast images below show the positions of the cells. Scale bar: 10 μm. ( C ) Fluorescence images as in A and B were thresholded and the resulting pixel intensity was divided by the cell area. The compiled quantified data are normalized to WT. Data were compiled from two experiments and 10 images each. P
    Rabbit Anti Laminin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 56 article reviews
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    99
    Millipore anti laminin antibody
    Muscle fibre type in COPD versus Control A , representative immunoflorescence images labelled for type I (blue), type IIa (red), type IIx (green) and <t>laminin</t> (green), where grouped fast fibres are indicated (*). Fibres co‐expressing type IIa and IIx were seen in all groups and these appear reddish/green/brown in the images (bar = 100 μm). B , fibre type proportions. C , frequency of fibres completely surrounded by fibres of the same type (Grouped Fibres). * P
    Anti Laminin Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti laminin antibody/product/Millipore
    Average 99 stars, based on 4388 article reviews
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    99
    Millipore rabbit anti laminin
    Glycosylation abnormalities of α-dystroglycan and recovery of glycosylation by co-injection of FKRP mRNA with MO2. ( A – F ) Immunostaining of 4 dpf fish co-injected FKRP mRNA with anti α-dystroglycan (VIA4-1). (A) Control MO2. (B) FKRP MO2 injected fish. (C) Human normal FKRP mRNA co-injected fish. (D) Human mutated FKRP mRNA (L276I) co-injected fish. (E) Human mutated FKRP mRNA (C318Y) co-injected fish. (F) Human mutated FKRP mRNA (A455D) co-injected fish. Bar: 100 µm. ( G ) <t>laminin</t> overlay assay and western blot with α- and β-dystroglycan antibody.
    Rabbit Anti Laminin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1577 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti laminin/product/Millipore
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    rabbit anti laminin - by Bioz Stars, 2020-09
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    99
    Abcam rabbit anti laminin
    Histology of mitral valve scaffolds. Representative histology analysis of fresh and decellularized (Decell) mitral valve leaflets and chordae stained with Movats' Pentachrome (A, F) , VVG in (B and E) , and immunohistochemistry for type IV collagen and <t>laminin</t> (C, D)
    Rabbit Anti Laminin, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 404 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti laminin/product/Abcam
    Average 99 stars, based on 404 article reviews
    Price from $9.99 to $1999.99
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    99
    Millipore rabbit polyclonal anti laminin
    Histology of mitral valve scaffolds. Representative histology analysis of fresh and decellularized (Decell) mitral valve leaflets and chordae stained with Movats' Pentachrome (A, F) , VVG in (B and E) , and immunohistochemistry for type IV collagen and <t>laminin</t> (C, D)
    Rabbit Polyclonal Anti Laminin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti laminin/product/Millipore
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    93
    Agilent technologies rabbit anti laminin
    Infiltration of human breast cancer tumors by mouse TAMs . ( a ) BT474-M1 and ( b ) HCC1954 tumors were established in CB17-SCID-beige mice. Tumors were harvested at 4 weeks after tumor cell injection and sections analyzed for vascularization using antibodies against mouse CD31 (endothelial marker); for tumor ECM, using antibodies against mouse <t>laminin;</t> for leukocyte infiltration using antibodies against mouse CD45; and for TAM infiltration using antibodies against mouse F4/80. Bar = 40 µm. TAM, tumor-associated macrophage.
    Rabbit Anti Laminin, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 214 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam rabbit anti human laminin
    Engineered microvasculature exhibits appropriate endothelial barrier function a) Schematic showing while seeded endothelial cells initially adhered to gelatin, they appropriately assembly basement membrane during culture. b) Representative 3D confocal microscopy immunostaining images of the adherens junction protein VE-cadherin and the basement membrane proteins <t>laminin</t> and collagen-IV, self-deposited by HUVECs after 14 days of culture. c) A representative stitched composite of epi-fluorescence images after 15-minute perfusion of BSA-AF594. d) Fluorescence intensity of the BSA-AF594 along the linescan across the engineered microvasculature in panel b. e) Higher magnification view of the engineered microvasculature demonstrating that the system is impermeable to BSA under physiologic flow conditions. f) In acellular (non-endothelialized) microsystems, significant diffusion of BSA occured as early as 5 minutes after perfusion (dashed lines define the microchannel borders). g) Representative plots of BSA-AF594 fluorescence intensity over time in permeability assays for both acellular and endothelialized microdevices. h) Quantified apparent permeability (P a ) of the engineered microvasculature to BSA remained similar across different endothelial cell types, and is approximately 40× less than that in acellular microchannels. Data was plotted as the mean ± s.d. with n=4 independent biological replicates expect for HLMVECs (n=3). P -values were calculated using one-way ANOVA with Bonferroni’s post hoc test (*** P
    Rabbit Anti Human Laminin, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Agilent technologies polyclonal rabbit anti laminin
    Engineered microvasculature exhibits appropriate endothelial barrier function a) Schematic showing while seeded endothelial cells initially adhered to gelatin, they appropriately assembly basement membrane during culture. b) Representative 3D confocal microscopy immunostaining images of the adherens junction protein VE-cadherin and the basement membrane proteins <t>laminin</t> and collagen-IV, self-deposited by HUVECs after 14 days of culture. c) A representative stitched composite of epi-fluorescence images after 15-minute perfusion of BSA-AF594. d) Fluorescence intensity of the BSA-AF594 along the linescan across the engineered microvasculature in panel b. e) Higher magnification view of the engineered microvasculature demonstrating that the system is impermeable to BSA under physiologic flow conditions. f) In acellular (non-endothelialized) microsystems, significant diffusion of BSA occured as early as 5 minutes after perfusion (dashed lines define the microchannel borders). g) Representative plots of BSA-AF594 fluorescence intensity over time in permeability assays for both acellular and endothelialized microdevices. h) Quantified apparent permeability (P a ) of the engineered microvasculature to BSA remained similar across different endothelial cell types, and is approximately 40× less than that in acellular microchannels. Data was plotted as the mean ± s.d. with n=4 independent biological replicates expect for HLMVECs (n=3). P -values were calculated using one-way ANOVA with Bonferroni’s post hoc test (*** P
    Polyclonal Rabbit Anti Laminin, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Laminin assembly is compromised in mammary epithelial cells lacking DG expression. ( A ) Primary mammary epithelial cells from control or ΔDG K14-Cre mice were incubated with 10 ng/ml FITC-laminin-111 overnight. DG-expressing cells (WT) showed efficient assembly of laminin, whereas DG-knockout cells (KO) showed little assembled laminin. The phase-contrast images below show the positions of the cells. ( B ) Cells derived from the immortalized DG-knockout cell line (MEpL), transfected with DG (WT) or a vector control (KO) were overlaid with 10 ng/ml FITC-laminin-111 overnight. WT cells show efficient assembly of laminin, whereas the knockout cells show no assembled laminin. The differential interference contrast images below show the positions of the cells. Scale bar: 10 μm. ( C ) Fluorescence images as in A and B were thresholded and the resulting pixel intensity was divided by the cell area. The compiled quantified data are normalized to WT. Data were compiled from two experiments and 10 images each. P

    Journal: Journal of Cell Science

    Article Title: Dystroglycan controls signaling of multiple hormones through modulation of STAT5 activity

    doi: 10.1242/jcs.070680

    Figure Lengend Snippet: Laminin assembly is compromised in mammary epithelial cells lacking DG expression. ( A ) Primary mammary epithelial cells from control or ΔDG K14-Cre mice were incubated with 10 ng/ml FITC-laminin-111 overnight. DG-expressing cells (WT) showed efficient assembly of laminin, whereas DG-knockout cells (KO) showed little assembled laminin. The phase-contrast images below show the positions of the cells. ( B ) Cells derived from the immortalized DG-knockout cell line (MEpL), transfected with DG (WT) or a vector control (KO) were overlaid with 10 ng/ml FITC-laminin-111 overnight. WT cells show efficient assembly of laminin, whereas the knockout cells show no assembled laminin. The differential interference contrast images below show the positions of the cells. Scale bar: 10 μm. ( C ) Fluorescence images as in A and B were thresholded and the resulting pixel intensity was divided by the cell area. The compiled quantified data are normalized to WT. Data were compiled from two experiments and 10 images each. P

    Article Snippet: Sections were then incubated overnight at 4°C with the following primary antibodies diluted in blocking buffer: 1:100 rabbit anti-ZO-1 (Zymed), 1:200 rabbit anti-laminin, 1:50 rabbit anti-P-STAT5 (Invitrogen), 1:50 rabbit anti-STAT5 (Santa Cruz Biotechnology), 1:1000 rabbit anti-keratin-14 (Sigma), 1:30 rat GoH3 mAb anti-α6-integrin (Chemicon), 1:50 mouse anti-β-DG (MANDAG-2, Developmental Studies Hybridoma Bank, Iowa), 1:400 mouse anti-α-smooth-muscle-actin (clone 1A4, Sigma), 1:200 mouse E-cadherin (BD Transduction Labs).

    Techniques: Expressing, Mouse Assay, Incubation, Knock-Out, Derivative Assay, Transfection, Plasmid Preparation, Fluorescence

    K14-Cre activity efficiently ablates DG expression. ( A ) Frozen sections of L1 stage mammary glands from control or ΔDG K14-Cre mice co-stained with anti-laminin (Ln) and anti-DG (DG) antibodies. The right panels show the merged laminin and DG co-staining. DG staining was absent from the ΔDG K14-Cre epithelium. Arrows indicate the green laminin staining the exterior of the gland, whereas arrowheads indicate red DG staining the basal surface of the epithelium. Areas of yellow reveal overlap of laminin with its receptor, DG. Scale bar: 20 μm. ( B ) Protein extracts of primary cell cultures from control and ΔDG K14-Cre glands immunoblotted with the antibodies indicated on the right. No β-DG was detected in mammary epithelial cells cultured from ΔDG K14-Cre mice.

    Journal: Journal of Cell Science

    Article Title: Dystroglycan controls signaling of multiple hormones through modulation of STAT5 activity

    doi: 10.1242/jcs.070680

    Figure Lengend Snippet: K14-Cre activity efficiently ablates DG expression. ( A ) Frozen sections of L1 stage mammary glands from control or ΔDG K14-Cre mice co-stained with anti-laminin (Ln) and anti-DG (DG) antibodies. The right panels show the merged laminin and DG co-staining. DG staining was absent from the ΔDG K14-Cre epithelium. Arrows indicate the green laminin staining the exterior of the gland, whereas arrowheads indicate red DG staining the basal surface of the epithelium. Areas of yellow reveal overlap of laminin with its receptor, DG. Scale bar: 20 μm. ( B ) Protein extracts of primary cell cultures from control and ΔDG K14-Cre glands immunoblotted with the antibodies indicated on the right. No β-DG was detected in mammary epithelial cells cultured from ΔDG K14-Cre mice.

    Article Snippet: Sections were then incubated overnight at 4°C with the following primary antibodies diluted in blocking buffer: 1:100 rabbit anti-ZO-1 (Zymed), 1:200 rabbit anti-laminin, 1:50 rabbit anti-P-STAT5 (Invitrogen), 1:50 rabbit anti-STAT5 (Santa Cruz Biotechnology), 1:1000 rabbit anti-keratin-14 (Sigma), 1:30 rat GoH3 mAb anti-α6-integrin (Chemicon), 1:50 mouse anti-β-DG (MANDAG-2, Developmental Studies Hybridoma Bank, Iowa), 1:400 mouse anti-α-smooth-muscle-actin (clone 1A4, Sigma), 1:200 mouse E-cadherin (BD Transduction Labs).

    Techniques: Activity Assay, Expressing, Mouse Assay, Staining, Cell Culture

    Laminin-111 enhances the expression of genes induced by growth hormone. MEpL cells expressing WT DG were overlaid with medium containing 100 μg/ml laminin-111 or 1.5% matrigel in the presence or absence of growth hormone for 72 hours. ( A ) Cell lysates were analyzed for the presence of the milk protein β-casein by western blot. E-cadherin was used as a loading control. The ratio of β-casein to E-cadherin was normalized to untreated cells and is indicated below the immunoblots. ( B ) Total RNA was extracted from these cells and RT-qPCR was performed for the expression of Igf1 and normalized to Gapdh expression. The data is expressed as the fold change of mRNA in hormone-treated samples versus control, untreated samples (* P

    Journal: Journal of Cell Science

    Article Title: Dystroglycan controls signaling of multiple hormones through modulation of STAT5 activity

    doi: 10.1242/jcs.070680

    Figure Lengend Snippet: Laminin-111 enhances the expression of genes induced by growth hormone. MEpL cells expressing WT DG were overlaid with medium containing 100 μg/ml laminin-111 or 1.5% matrigel in the presence or absence of growth hormone for 72 hours. ( A ) Cell lysates were analyzed for the presence of the milk protein β-casein by western blot. E-cadherin was used as a loading control. The ratio of β-casein to E-cadherin was normalized to untreated cells and is indicated below the immunoblots. ( B ) Total RNA was extracted from these cells and RT-qPCR was performed for the expression of Igf1 and normalized to Gapdh expression. The data is expressed as the fold change of mRNA in hormone-treated samples versus control, untreated samples (* P

    Article Snippet: Sections were then incubated overnight at 4°C with the following primary antibodies diluted in blocking buffer: 1:100 rabbit anti-ZO-1 (Zymed), 1:200 rabbit anti-laminin, 1:50 rabbit anti-P-STAT5 (Invitrogen), 1:50 rabbit anti-STAT5 (Santa Cruz Biotechnology), 1:1000 rabbit anti-keratin-14 (Sigma), 1:30 rat GoH3 mAb anti-α6-integrin (Chemicon), 1:50 mouse anti-β-DG (MANDAG-2, Developmental Studies Hybridoma Bank, Iowa), 1:400 mouse anti-α-smooth-muscle-actin (clone 1A4, Sigma), 1:200 mouse E-cadherin (BD Transduction Labs).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    Muscle fibre type in COPD versus Control A , representative immunoflorescence images labelled for type I (blue), type IIa (red), type IIx (green) and laminin (green), where grouped fast fibres are indicated (*). Fibres co‐expressing type IIa and IIx were seen in all groups and these appear reddish/green/brown in the images (bar = 100 μm). B , fibre type proportions. C , frequency of fibres completely surrounded by fibres of the same type (Grouped Fibres). * P

    Journal: The Journal of Physiology

    Article Title: Smoke‐induced neuromuscular junction degeneration precedes the fibre type shift and atrophy in chronic obstructive pulmonary disease

    doi: 10.1113/JP275558

    Figure Lengend Snippet: Muscle fibre type in COPD versus Control A , representative immunoflorescence images labelled for type I (blue), type IIa (red), type IIx (green) and laminin (green), where grouped fast fibres are indicated (*). Fibres co‐expressing type IIa and IIx were seen in all groups and these appear reddish/green/brown in the images (bar = 100 μm). B , fibre type proportions. C , frequency of fibres completely surrounded by fibres of the same type (Grouped Fibres). * P

    Article Snippet: A cocktail of primary antibodies, myosin heavy chain (MHC) Type I (BA‐F8, DSHB, University of Iowa), IIa (Sc71, DSHB), IIx (6H1 DSHB) and laminin (L9393, Sigma, St Louis, MO, USA), was applied for 1 h at room temperature, followed by a series of PBS washes and incubation with fluorescence‐conjugated secondary antibodies for 1 h at room temperature.

    Techniques: Expressing

    Peripheral nerve endings are altered in Gars C201R /+ mice. (A) Representative SV2/2H3 + (green) muscle spindles from wild-type (left) and Gars C201R /+ (right) soleus muscles. Anti-laminin highlights the muscle basement membrane (red). N.B., the lack of SV2/3H3 positivity surrounding the central nuclei (DAPI, blue) of the mutant spindle (arrows). Images are single confocal planes. (B-C) Gars C201R /+ mice have significantly fewer spindles per soleus muscle (B, ** P = 0.005, unpaired t -test). Furthermore, mutant spindles display significant denervation (C, *** P

    Journal: bioRxiv

    Article Title: Sensory neuron fate is developmentally perturbed by Gars mutations causing human neuropathy

    doi: 10.1101/071159

    Figure Lengend Snippet: Peripheral nerve endings are altered in Gars C201R /+ mice. (A) Representative SV2/2H3 + (green) muscle spindles from wild-type (left) and Gars C201R /+ (right) soleus muscles. Anti-laminin highlights the muscle basement membrane (red). N.B., the lack of SV2/3H3 positivity surrounding the central nuclei (DAPI, blue) of the mutant spindle (arrows). Images are single confocal planes. (B-C) Gars C201R /+ mice have significantly fewer spindles per soleus muscle (B, ** P = 0.005, unpaired t -test). Furthermore, mutant spindles display significant denervation (C, *** P

    Article Snippet: The following primary antibodies were used ( - ): sheep anti-CGRP (1/200, Enzo Life Sciences, Farmingdale, NY, BML-CA1137), rabbit anti-cleaved-caspase 3 (1/500, Cell Signalling Technology, Danvers, MA, 9661), rabbit anti-laminin (1/1000, Sigma, L9393), mouse anti-neurofilament (2H3, 1/50 or 1/250, developed by Thomas M. Jessell and Jane Dodd, Developmental Studies Hybridoma Bank, Iowa City, IA, supernatant), mouse anti-NF200 (1/500, Sigma, N0142), rabbit anti-parvalbumin (1/1000, Swant, Marly, Switzerland, PV27), rabbit anti-peripherin (1/500, Merck Millipore, AB1530), rabbit anti-PGP9.5 (1/1000, UltraClone, Isle of White, UK, 31A3), rabbit anti-PSD95 (1/200, Frontier Institute, Ishikari, Japan, Af628), mouse pan anti-synaptic vesicle 2 (SV2, 1/250, developed by Kathleen M. Buckley, Developmental Studies Hybridoma Bank, Iowa City, IA, concentrate), guinea pig anti-synaptophysin (1/200, Frontier Institute, Af300), chicken anti-β III-tubulin (1/500, Abcam, Cambridge, UK, ab41489), and mouse anti-β III-tubulin (1/500, Covance, Princeton, NJ, mms-435P).

    Techniques: Mouse Assay, Mutagenesis

    Differential cellular characteristics of the neural tube at neural crest ( NC ) and roof plate ( RP ) stages. a , b Transverse sections of the flank region of E4.5 avian embryos whose hemi-neural tubes ( NTs ) were electroporated with a control GFP plasmid at E2 ( a ) or E3.5 ( b ). Note the contribution of labeled cells to NC derivatives including melanocytes ( arrow in a) following early but not late stage electroporations. c , d Bromodeoxyuridine ( BrdU ) incorporation following a 1-h pulse at NC (E2–E2.5, c) or RP (E3.5, d) stages. Dashed lines in insets mark the dorsal NT domain that was quantified (see text for details). Note the presence of the BrdU+ nuclei (Red) in c – c” but not in the equivalent dashed area in d – d” . Nuclei were visualized with Hoechst. e – j Antibody staining for epithelial (ZO-1, N-cadherin, laminin) or ciliary (Arl13b) markers. Arrows point to disorganized cilia ( e’ ), the absence of N-cadherin in the dorsal NT compared to more ventral regions ( g ), and an incomplete laminin-containing basal lamina ( i , i’ ) at the NC stage. In contrast, note the apically oriented cilia ( f ), positive N-cadherin immunostaining ( h ), and continuous laminin expression ( j ) in the dorsal NT at the RP stage ( arrowheads in f , h , and j’ ). Ect ectoderm. Bar in a , b , d , h , j = 80 μM; c = 50 μM; c’ , d’ , e = 30 μM; f , g , i’ = 40 μM; e’ =15 μM; j = 240 μM; i =140 μM

    Journal: BMC Biology

    Article Title: Dynamics of BMP and Hes1/Hairy1 signaling in the dorsal neural tube underlies the transition from neural crest to definitive roof plate

    doi: 10.1186/s12915-016-0245-6

    Figure Lengend Snippet: Differential cellular characteristics of the neural tube at neural crest ( NC ) and roof plate ( RP ) stages. a , b Transverse sections of the flank region of E4.5 avian embryos whose hemi-neural tubes ( NTs ) were electroporated with a control GFP plasmid at E2 ( a ) or E3.5 ( b ). Note the contribution of labeled cells to NC derivatives including melanocytes ( arrow in a) following early but not late stage electroporations. c , d Bromodeoxyuridine ( BrdU ) incorporation following a 1-h pulse at NC (E2–E2.5, c) or RP (E3.5, d) stages. Dashed lines in insets mark the dorsal NT domain that was quantified (see text for details). Note the presence of the BrdU+ nuclei (Red) in c – c” but not in the equivalent dashed area in d – d” . Nuclei were visualized with Hoechst. e – j Antibody staining for epithelial (ZO-1, N-cadherin, laminin) or ciliary (Arl13b) markers. Arrows point to disorganized cilia ( e’ ), the absence of N-cadherin in the dorsal NT compared to more ventral regions ( g ), and an incomplete laminin-containing basal lamina ( i , i’ ) at the NC stage. In contrast, note the apically oriented cilia ( f ), positive N-cadherin immunostaining ( h ), and continuous laminin expression ( j ) in the dorsal NT at the RP stage ( arrowheads in f , h , and j’ ). Ect ectoderm. Bar in a , b , d , h , j = 80 μM; c = 50 μM; c’ , d’ , e = 30 μM; f , g , i’ = 40 μM; e’ =15 μM; j = 240 μM; i =140 μM

    Article Snippet: Immunohistochemistry and in situ hybridization Antibodies against HNK1 (CD57, BD Biosciences, San Jose, CA, USA Cat#559048, 1:500), Arl13b (from Tamara Caspary, 1:1000), ZO-1 (Thermo Fisher Scientific, Waltham, MA USA, cat#402200, 1:100), N-cadherin (R & D Systems, Minneapolis, MN, USA., cat#BTA7, 5 μg/ml), laminin (Sigma-Aldrich, St. St. Louis, MO, USA cat#L9393,1:100), BrdU (G3G4, Developmental Studies Hybridoma bank, Iowa City, Iowa, USA 1:100), and phosphorylated Smad 1-5-8 (pSMAD, from Ed Laufer, 1:1000) were used as previously described [ ].

    Techniques: Plasmid Preparation, Labeling, BrdU Incorporation Assay, Staining, Immunostaining, Expressing

    Muscle fibre atrophy is independent of fibre type in lung cancer cachexia . Cross-sectional of individual muscle fibres was assessed using immunohistochemical staining of laminin. Assessment of fibres expressing different myosin heavy chain isoforms was performed using immunohistochemistry and myosin adenosine 5′-triphosphatase staining.

    Journal: Journal of Cachexia, Sarcopenia and Muscle

    Article Title: Preserved muscle oxidative metabolic phenotype in newly diagnosed non-small cell lung cancer cachexia

    doi: 10.1002/jcsm.12007

    Figure Lengend Snippet: Muscle fibre atrophy is independent of fibre type in lung cancer cachexia . Cross-sectional of individual muscle fibres was assessed using immunohistochemical staining of laminin. Assessment of fibres expressing different myosin heavy chain isoforms was performed using immunohistochemistry and myosin adenosine 5′-triphosphatase staining.

    Article Snippet: Slides were incubated with primary: anti-laminin (dilution 1:50; #L-9393, Sigma, St. Louis, MO, USA); anti-type I MyHC (dilution 1:40; #A4840, Developmental Studies Hybridoma Bank (DSHB), Iowa City, IA, USA); and anti-type II MyHC (dilution 1:40; #N2261, Santa Cruz, CA, USA), and secondary antibodies: Alexa Fluor 350 (dilution 1:100; #A-11069); Alexa Fluor 488 (dilution 1:1000; #A-21121) and Alexa Fluor 555 (dilution 1:1000; #A-21426, Invitrogen, Madison, Wisconsin, USA).

    Techniques: Immunohistochemistry, Staining, Expressing

    Imaging of structural details of sporadically labeled podocytes in uninjured Coll1α1GCE + ;R26Tomato + kidneys by conventional fluorescence microscopy. (A–C) Whole antilaminin-stained glomerulus (green) highlighting close association of tdTomato+ network of podocyte processes and laminin+ glomerular basement membrane. Several regions of interest are illustrated. (D) A region with interdigitating foot processes from two neighboring podocytes that are both fate-labeled is shown. Of note, resolution is insufficient to delineate details beyond the nucleated cell bodies (*) and main processes (α). (E–G) By contrast, these cutouts highlight areas in which only every other foot process is genetically labeled, and neighboring pedicles remain optically silent. Resolution is adequate to assess morphology and extension of single tertiary foot processes. Note that the lateral branching of tertiary foot processes out from main processes resembles the typical arrangement of fish bones. (H–K) Antilaminin-stained glomerular capillary loop (green) covered by a network of genetically labeled podocyte foot processes (red). (K) The three-dimensional view illustrates how foot processes conform to the ridge-like elevation of the underlying capillary (top to bottom=2.2 μm). (L) Example of a labeled podocyte with classic podocyte architecture: a nucleated cell body (*) and a primary process (α) branching into several secondary processes (β), which again branch into many minor, tertiary foot processes at regular intervals. (M) Example of tertiary foot process branching in very close proximity to the nucleated cell body (*). Scale bars, 10 μm in A–C; 2 μm in D–M.

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: Imaging of Podocyte Foot Processes by Fluorescence Microscopy

    doi: 10.1681/ASN.2011100988

    Figure Lengend Snippet: Imaging of structural details of sporadically labeled podocytes in uninjured Coll1α1GCE + ;R26Tomato + kidneys by conventional fluorescence microscopy. (A–C) Whole antilaminin-stained glomerulus (green) highlighting close association of tdTomato+ network of podocyte processes and laminin+ glomerular basement membrane. Several regions of interest are illustrated. (D) A region with interdigitating foot processes from two neighboring podocytes that are both fate-labeled is shown. Of note, resolution is insufficient to delineate details beyond the nucleated cell bodies (*) and main processes (α). (E–G) By contrast, these cutouts highlight areas in which only every other foot process is genetically labeled, and neighboring pedicles remain optically silent. Resolution is adequate to assess morphology and extension of single tertiary foot processes. Note that the lateral branching of tertiary foot processes out from main processes resembles the typical arrangement of fish bones. (H–K) Antilaminin-stained glomerular capillary loop (green) covered by a network of genetically labeled podocyte foot processes (red). (K) The three-dimensional view illustrates how foot processes conform to the ridge-like elevation of the underlying capillary (top to bottom=2.2 μm). (L) Example of a labeled podocyte with classic podocyte architecture: a nucleated cell body (*) and a primary process (α) branching into several secondary processes (β), which again branch into many minor, tertiary foot processes at regular intervals. (M) Example of tertiary foot process branching in very close proximity to the nucleated cell body (*). Scale bars, 10 μm in A–C; 2 μm in D–M.

    Article Snippet: The following primary antibodies were used for immunostaining: guinea pig antinephrin (1:100; Progen Biotechnik), rat anti-CD31 (1:100; eBioscience), rat anti-PDGFRβ (1:200; eBioscience), and rabbit antilaminin (1:500; Sigma).

    Techniques: Imaging, Labeling, Fluorescence, Microscopy, Staining, Fluorescence In Situ Hybridization

    Administration of tamoxifen/4-OHT to bigenic Coll1α1GCE + ;reporter + mice fate-labels podocytes in a dose-dependent fashion. (A) Experimental approach for conditional labeling of podocytes in vivo . Injection of tamoxifen or its active metabolite 4-OHT into bigenic Coll1α1GCE + ;R26LacZ + , Coll1α1GCE + ;mT/mG + , or Coll1α1GCE + ;R26Tomato + mice enables temporary translocation of the GFP-Cre-ERT2 fusion protein into the cell nucleus, removing the LoxP -flanked DNA sequence of the reporter allele and leading to permanent expression of LacZ, membrane-targeted EGFP, and tdTomato, respectively. (B–D) Validation of the labeling strategy. Only in adult kidneys of tamoxifen/4-OHT–injected bigenic animals is there robust visceral epithelial expression of LacZ, membrane-targeted EGFP, or tdTomato. Insets (D, right) show a labeled pericyte (red) in the medulla of a Coll1α1GCE + ;R26Tomato + kidney, and antilaminin staining (green) highlights its tubulointerstitial localization. Scale bars, 50 μm. (E–G) Semiquantitative assessment of recombination efficiency in glomeruli in relation to time point and dose of tamoxifen/4-OHT administration. Animals injected only one time at P0 exhibited no evidence of recombination in glomeruli of the outer cortex, suggesting a lack of collagen1α1 promoter activity in podocyte progenitors of the nephrogenic zone at the time point of injection. Single injection at P3 resulted in a more homogenous but mostly partial labeling of glomeruli throughout the cortex. By contrast, repeated tamoxifen/4-OHT injections (P0 and P3) labeled the majority of podocytes in almost all glomeruli. Quantitation was performed in Coll1α1GCE + ;mT/mG + and Coll1α1GCE + ;R26Tomato + mice, respectively. Data are given as mean ± SEM; n =3–4 for each data point.

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: Imaging of Podocyte Foot Processes by Fluorescence Microscopy

    doi: 10.1681/ASN.2011100988

    Figure Lengend Snippet: Administration of tamoxifen/4-OHT to bigenic Coll1α1GCE + ;reporter + mice fate-labels podocytes in a dose-dependent fashion. (A) Experimental approach for conditional labeling of podocytes in vivo . Injection of tamoxifen or its active metabolite 4-OHT into bigenic Coll1α1GCE + ;R26LacZ + , Coll1α1GCE + ;mT/mG + , or Coll1α1GCE + ;R26Tomato + mice enables temporary translocation of the GFP-Cre-ERT2 fusion protein into the cell nucleus, removing the LoxP -flanked DNA sequence of the reporter allele and leading to permanent expression of LacZ, membrane-targeted EGFP, and tdTomato, respectively. (B–D) Validation of the labeling strategy. Only in adult kidneys of tamoxifen/4-OHT–injected bigenic animals is there robust visceral epithelial expression of LacZ, membrane-targeted EGFP, or tdTomato. Insets (D, right) show a labeled pericyte (red) in the medulla of a Coll1α1GCE + ;R26Tomato + kidney, and antilaminin staining (green) highlights its tubulointerstitial localization. Scale bars, 50 μm. (E–G) Semiquantitative assessment of recombination efficiency in glomeruli in relation to time point and dose of tamoxifen/4-OHT administration. Animals injected only one time at P0 exhibited no evidence of recombination in glomeruli of the outer cortex, suggesting a lack of collagen1α1 promoter activity in podocyte progenitors of the nephrogenic zone at the time point of injection. Single injection at P3 resulted in a more homogenous but mostly partial labeling of glomeruli throughout the cortex. By contrast, repeated tamoxifen/4-OHT injections (P0 and P3) labeled the majority of podocytes in almost all glomeruli. Quantitation was performed in Coll1α1GCE + ;mT/mG + and Coll1α1GCE + ;R26Tomato + mice, respectively. Data are given as mean ± SEM; n =3–4 for each data point.

    Article Snippet: The following primary antibodies were used for immunostaining: guinea pig antinephrin (1:100; Progen Biotechnik), rat anti-CD31 (1:100; eBioscience), rat anti-PDGFRβ (1:200; eBioscience), and rabbit antilaminin (1:500; Sigma).

    Techniques: Mouse Assay, Labeling, In Vivo, Injection, Translocation Assay, Sequencing, Expressing, Staining, Activity Assay, Quantitation Assay

    Glycosylation abnormalities of α-dystroglycan and recovery of glycosylation by co-injection of FKRP mRNA with MO2. ( A – F ) Immunostaining of 4 dpf fish co-injected FKRP mRNA with anti α-dystroglycan (VIA4-1). (A) Control MO2. (B) FKRP MO2 injected fish. (C) Human normal FKRP mRNA co-injected fish. (D) Human mutated FKRP mRNA (L276I) co-injected fish. (E) Human mutated FKRP mRNA (C318Y) co-injected fish. (F) Human mutated FKRP mRNA (A455D) co-injected fish. Bar: 100 µm. ( G ) laminin overlay assay and western blot with α- and β-dystroglycan antibody.

    Journal: Human Molecular Genetics

    Article Title: Zebrafish models for human FKRP muscular dystrophies

    doi: 10.1093/hmg/ddp528

    Figure Lengend Snippet: Glycosylation abnormalities of α-dystroglycan and recovery of glycosylation by co-injection of FKRP mRNA with MO2. ( A – F ) Immunostaining of 4 dpf fish co-injected FKRP mRNA with anti α-dystroglycan (VIA4-1). (A) Control MO2. (B) FKRP MO2 injected fish. (C) Human normal FKRP mRNA co-injected fish. (D) Human mutated FKRP mRNA (L276I) co-injected fish. (E) Human mutated FKRP mRNA (C318Y) co-injected fish. (F) Human mutated FKRP mRNA (A455D) co-injected fish. Bar: 100 µm. ( G ) laminin overlay assay and western blot with α- and β-dystroglycan antibody.

    Article Snippet: Membranes were incubated with rabbit anti-laminin (1:5000, Sigma) followed by anti-rabbit IgG-HRP (1:15000, Invitrogen).

    Techniques: Injection, Immunostaining, Fluorescence In Situ Hybridization, Overlay Assay, Western Blot

    The gaps between myofibers can be repaired by co-injection of normal FKRP mRNA with MO2 but not all human mutant alleles. Co-immunostaining of 4 dpf fish co-injected FKRP mRNA with anti MHC (green) and anti laminin (red). ( A ) Control MO2. ( B ) Human normal FKRP mRNA co-injected fish. ( C ) FKRP MO2 injected fish. ( D ) Human mutated FKRP mRNA (L276I) co-injected fish. ( E ) Human mutated FKRP mRNA (C318Y) co-injected fish. ( F ) Human mutated FKRP mRNA (A455D) co-injected fish. Bar: 50 µm. In control and human control mRNA co-injected fish, the myofibers are adjacent at myosepta (arrows in A and B); however, FKRP morphant has the gap between myosepta (arrowhead in C). In L276I mutant fish, the gap is also narrowed (arrow in D). In C318Y and A455D mutant fish, there still remain gaps similar to that found in FKRP morphant fish (arrows in E and F).

    Journal: Human Molecular Genetics

    Article Title: Zebrafish models for human FKRP muscular dystrophies

    doi: 10.1093/hmg/ddp528

    Figure Lengend Snippet: The gaps between myofibers can be repaired by co-injection of normal FKRP mRNA with MO2 but not all human mutant alleles. Co-immunostaining of 4 dpf fish co-injected FKRP mRNA with anti MHC (green) and anti laminin (red). ( A ) Control MO2. ( B ) Human normal FKRP mRNA co-injected fish. ( C ) FKRP MO2 injected fish. ( D ) Human mutated FKRP mRNA (L276I) co-injected fish. ( E ) Human mutated FKRP mRNA (C318Y) co-injected fish. ( F ) Human mutated FKRP mRNA (A455D) co-injected fish. Bar: 50 µm. In control and human control mRNA co-injected fish, the myofibers are adjacent at myosepta (arrows in A and B); however, FKRP morphant has the gap between myosepta (arrowhead in C). In L276I mutant fish, the gap is also narrowed (arrow in D). In C318Y and A455D mutant fish, there still remain gaps similar to that found in FKRP morphant fish (arrows in E and F).

    Article Snippet: Membranes were incubated with rabbit anti-laminin (1:5000, Sigma) followed by anti-rabbit IgG-HRP (1:15000, Invitrogen).

    Techniques: Injection, Mutagenesis, Immunostaining, Fluorescence In Situ Hybridization

    Indirect immunofluorescence microscopy for N-cadherin in glomeruli of quail (A–D’), chicken (E–F’). N-cadherin (A–F’) is stained green; podocin (B’, E’) and laminin (C’, D’, F’) are red in double-labeled immunofluorescence. N-cadherin is seen along the glomerular capillary wall in both birds. At least two types of staining pattern, weakly diffuse (arrows in D, D’, F, F’) and distinctly granular (arrowheads in D, D’, F, F’), are observed. (D) and (D’) are enlargements of the boxed areas in (C) and (C’), respectively. N-cadherin colocalizes with podocin (B, B’, E, E’). Scale (A) 100 µm; (B’, C’, E’) 10 µm; (D’, F’) 5 µm.

    Journal: Journal of Histochemistry and Cytochemistry

    Article Title: Avian Podocytes, Which Lack Nephrin, Use Adherens Junction Proteins at Intercellular Junctions

    doi: 10.1369/0022155415611708

    Figure Lengend Snippet: Indirect immunofluorescence microscopy for N-cadherin in glomeruli of quail (A–D’), chicken (E–F’). N-cadherin (A–F’) is stained green; podocin (B’, E’) and laminin (C’, D’, F’) are red in double-labeled immunofluorescence. N-cadherin is seen along the glomerular capillary wall in both birds. At least two types of staining pattern, weakly diffuse (arrows in D, D’, F, F’) and distinctly granular (arrowheads in D, D’, F, F’), are observed. (D) and (D’) are enlargements of the boxed areas in (C) and (C’), respectively. N-cadherin colocalizes with podocin (B, B’, E, E’). Scale (A) 100 µm; (B’, C’, E’) 10 µm; (D’, F’) 5 µm.

    Article Snippet: Rabbit anti-laminin antibody (Sigma-Aldrich) was raised against laminin from EHS mouse sarcoma and reacted with laminin of human, mammal, avian, reptilian and amphibian sources.

    Techniques: Immunofluorescence, Microscopy, Staining, Labeling

    Indirect immunofluorescence microscopy of quail embryonic kidney for N-cadherin. N-cadherin is stained green (A–G); podocin (A, B) and laminin (C-G) are stained red. Podocin staining shows N-cadherin in nephrogenic zones near the surface of the kidney in addition to podocin-positive glomeruli and cells lining the surface of the kidney (A, B). Condensation (asterisk) around the tip of the ureteric bud is positive for N-cadherin (C). With maturation of glomeruli, N-cadherin staining is localized to podocytes (D–G). Scale (A) 100 µm; (B) 50 µm, (C–G) 20 µm.

    Journal: Journal of Histochemistry and Cytochemistry

    Article Title: Avian Podocytes, Which Lack Nephrin, Use Adherens Junction Proteins at Intercellular Junctions

    doi: 10.1369/0022155415611708

    Figure Lengend Snippet: Indirect immunofluorescence microscopy of quail embryonic kidney for N-cadherin. N-cadherin is stained green (A–G); podocin (A, B) and laminin (C-G) are stained red. Podocin staining shows N-cadherin in nephrogenic zones near the surface of the kidney in addition to podocin-positive glomeruli and cells lining the surface of the kidney (A, B). Condensation (asterisk) around the tip of the ureteric bud is positive for N-cadherin (C). With maturation of glomeruli, N-cadherin staining is localized to podocytes (D–G). Scale (A) 100 µm; (B) 50 µm, (C–G) 20 µm.

    Article Snippet: Rabbit anti-laminin antibody (Sigma-Aldrich) was raised against laminin from EHS mouse sarcoma and reacted with laminin of human, mammal, avian, reptilian and amphibian sources.

    Techniques: Immunofluorescence, Microscopy, Staining

    Expression of MyoD K133R in normal and fasted muscle fibres induces hypertrophy. A/ Representative cross-sectional areas of electroporated TA muscles from fed and starved mice. Adult Tibialis anterior (TA) muscles were electroporated with either a control (GFP), a MyoDwt-GFP or a MyoD mutant K133R-GFP plasmid. Mice (control or fasted by 2 days) were sacrificed 14 days later. Transfected fibres were identified on the basis of their GFP expression. The distribution of cross-sectional areas of EGFP-expressing fibres taken from four muscles under each conditions Fibres membranes were stained with polyclonal anti-laminin antibody. Scale bar, 20 µm. B/. Mean cross-sectional area of TA fibres in control (fed) and fasted mice (atrophy)±s.e.m. * P

    Journal: PLoS ONE

    Article Title: Inhibition of Atrogin-1/MAFbx Mediated MyoD Proteolysis Prevents Skeletal Muscle Atrophy In Vivo

    doi: 10.1371/journal.pone.0004973

    Figure Lengend Snippet: Expression of MyoD K133R in normal and fasted muscle fibres induces hypertrophy. A/ Representative cross-sectional areas of electroporated TA muscles from fed and starved mice. Adult Tibialis anterior (TA) muscles were electroporated with either a control (GFP), a MyoDwt-GFP or a MyoD mutant K133R-GFP plasmid. Mice (control or fasted by 2 days) were sacrificed 14 days later. Transfected fibres were identified on the basis of their GFP expression. The distribution of cross-sectional areas of EGFP-expressing fibres taken from four muscles under each conditions Fibres membranes were stained with polyclonal anti-laminin antibody. Scale bar, 20 µm. B/. Mean cross-sectional area of TA fibres in control (fed) and fasted mice (atrophy)±s.e.m. * P

    Article Snippet: Rabbit polyclonal anti-Laminin antibody (Sigma) was applied overnight to the treated sections.

    Techniques: Expressing, Mouse Assay, Mutagenesis, Plasmid Preparation, Transfection, Staining

    Comparison of the mouse and rat SEZ. (A, B) Coronal sections taken from normal adult mouse (A) and rat (B) brains, triple-immunostained for PH3 (in green , to mark mitotic cells, indicated by white arrows ), GFAP (in red , to mark astrocytes) and laminin

    Journal: Stem Cells and Development

    Article Title: The Number of Stem Cells in the Subependymal Zone of the Adult Rodent Brain is Correlated with the Number of Ependymal Cells and Not with the Volume of the Niche

    doi: 10.1089/scd.2011.0130

    Figure Lengend Snippet: Comparison of the mouse and rat SEZ. (A, B) Coronal sections taken from normal adult mouse (A) and rat (B) brains, triple-immunostained for PH3 (in green , to mark mitotic cells, indicated by white arrows ), GFAP (in red , to mark astrocytes) and laminin

    Article Snippet: Vibratome-cut (Leica) 70 μm thick brain sections were processed for immunohistochemistry using the rabbit antiphosphohistone 3 (anti-PH3; 1/500; Millipore), mouse anti-glial fibrillary acidic protein (anti-GFAP; 1/500; Sigma), rabbit anti-pan Laminin (1/200; Sigma) and rabbit anti-S100β (prediluted; Dako) antibodies, followed by incubation in appropriate Alexa-conjugated secondary antibodies (Invitrogen).

    Techniques:

    Laminin erosion fluorescence analysis of SNF2/L and SNF2-AP-S-L substrates immunostained with anti-laminin antibody at Day 0, Day 12 and Day 18 ( n = 10).

    Journal: Nanomaterials

    Article Title: The Effect of Laminin Surface Modification of Electrospun Silica Nanofiber Substrate on Neuronal Tissue Engineering

    doi: 10.3390/nano8030165

    Figure Lengend Snippet: Laminin erosion fluorescence analysis of SNF2/L and SNF2-AP-S-L substrates immunostained with anti-laminin antibody at Day 0, Day 12 and Day 18 ( n = 10).

    Article Snippet: The samples were then immersed in the rabbit anti-laminin primary antibody (diluted at 1:500, Sigma).

    Techniques: Fluorescence

    Fluorescence images of SNF2/L ( A – C ) and SNF2-AP-S-L ( D – F ) substrates immunostained with anti-laminin antibody at different time point: ( A , D ) Day 0, ( B , E ) at Day 12, ( C , F ) at Day 18. Scale bars represent 20 μm.

    Journal: Nanomaterials

    Article Title: The Effect of Laminin Surface Modification of Electrospun Silica Nanofiber Substrate on Neuronal Tissue Engineering

    doi: 10.3390/nano8030165

    Figure Lengend Snippet: Fluorescence images of SNF2/L ( A – C ) and SNF2-AP-S-L ( D – F ) substrates immunostained with anti-laminin antibody at different time point: ( A , D ) Day 0, ( B , E ) at Day 12, ( C , F ) at Day 18. Scale bars represent 20 μm.

    Article Snippet: The samples were then immersed in the rabbit anti-laminin primary antibody (diluted at 1:500, Sigma).

    Techniques: Fluorescence

    Histology of mitral valve scaffolds. Representative histology analysis of fresh and decellularized (Decell) mitral valve leaflets and chordae stained with Movats' Pentachrome (A, F) , VVG in (B and E) , and immunohistochemistry for type IV collagen and laminin (C, D)

    Journal: Tissue Engineering. Part A

    Article Title: Stabilized Collagen and Elastin-Based Scaffolds for Mitral Valve Tissue Engineering

    doi: 10.1089/ten.tea.2016.0032

    Figure Lengend Snippet: Histology of mitral valve scaffolds. Representative histology analysis of fresh and decellularized (Decell) mitral valve leaflets and chordae stained with Movats' Pentachrome (A, F) , VVG in (B and E) , and immunohistochemistry for type IV collagen and laminin (C, D)

    Article Snippet: The following antibodies were used: rabbit anti-Collagen IV (Ab6586; Abcam), rabbit anti-Laminin (Ab11575; Abcam), rabbit anti-Vimentin (Ab92547; Abcam), rabbit anti-α-Smooth Muscle Actin (Ab5694; Abcam), and rabbit anti-Integrin β1 (Ab52971; Abcam).

    Techniques: Staining, Immunohistochemistry

    Cellular and Morphological Evaluation of Pre-Innervated Constructs at Acute Time Point Following Implantation in a VML Model. a-d ) Longitudinal sections near the repair site of animals implanted with nanofibers with motor neurons + myocytes (MN-MYO). The nanofiber sheets were coated with Laminin prior to culturing cells and hence the stacked sheets were identified based on Laminin stain (red). Scale: 500µm. a′-d′) Magnified view of the region inside the white box. Thick bundles of myocytes (Phalloidin: green) and motor axons (NF-200: purple) were observed within the stacked nanofiber sheets. Scale: 100µm.

    Journal: bioRxiv

    Article Title: Pre-Innervated Tissue Engineered Muscle Promotes a Pro-Regenerative Microenvironment Following Volumetric Muscle Loss

    doi: 10.1101/840124

    Figure Lengend Snippet: Cellular and Morphological Evaluation of Pre-Innervated Constructs at Acute Time Point Following Implantation in a VML Model. a-d ) Longitudinal sections near the repair site of animals implanted with nanofibers with motor neurons + myocytes (MN-MYO). The nanofiber sheets were coated with Laminin prior to culturing cells and hence the stacked sheets were identified based on Laminin stain (red). Scale: 500µm. a′-d′) Magnified view of the region inside the white box. Thick bundles of myocytes (Phalloidin: green) and motor axons (NF-200: purple) were observed within the stacked nanofiber sheets. Scale: 100µm.

    Article Snippet: For staining of laminin, samples were incubated with rabbit-anti-laminin (1:500, abcam, ab11575) overnight at 4°C followed by AlexaFluor-568 antibody (1:500) for two hours at room temperature.

    Techniques: Construct, Staining

    BPMO-M23D and BPMO-M23D MSTN Administration Induce Substantial Body-wide Dystrophin Restoration (A) Western blot analysis showing dystrophin expression (dys + ) in DIA, GAS, SOL, TA, and heart muscles of BPMO-M23D- and BPMO-M23D MSTN-treated mdx mice. Each lane represents a sample from an individual mouse. Alpha-tubulin (α-tub + ) was used as an internal loading control for western blot. (B) Quantification of dystrophin expression by densitometric analysis of western blot. Following western blot evaluation, the intensity of dys + patterns was scored and normalized to the intensity of corresponding α-tub + patterns, and subsequently it was quantified based on a standard curve of C57 dystrophin. The results were expressed as the percentage of muscle type-matched C57 value (considered as 100%). Data are shown for individual muscle types or as an average of all types. (C and D) Immunostaining detecting dystrophin and laminin expression in treated muscles. Representative images of (C) DIA and (D) TA muscle sections for each group of mice are shown, respectively. Dystrophin-positive fibers were stained in green while laminin-positive fibers were stained in red. Nuclei were stained in blue with DAPI. Scale bars, 100 μm. (E) Quantification of dystrophin intensity levels in DIA, EDL, GAS, SOL, TA, and heart muscles. Following immunostaining for dystrophin, the mean dystrophin intensity was scored by ZEN software and normalized to the mean intensity of laminin detected on the same section. Results were expressed as the percentage of C57 value, considered as 100%. (F and G) Quantification of dystrophin-positive fibers was focused on (F) DIA and (G) TA muscles. The number of dystrophin- and laminin-positive fibers from five random fields of mid-belly muscle sections was counted. Only fibers showing continuous staining of dystrophin along the entire sarcolemma were considered as dystrophin positive and evaluated as the percentage of the number of total fibers (laminin positive) within the same image field. Results were expressed as the percentage of muscle type-matched C57 value, obtained in the same way and considered as 100%. Data in (B) and (E)–(G) are shown as means ± SEM; error bars represent the SEM; n = 10/group. Statistical comparison was two-tailed Student’s t test (*p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Systemic Antisense Therapeutics for Dystrophin and Myostatin Exon Splice Modulation Improve Muscle Pathology of Adult mdx Mice

    doi: 10.1016/j.omtn.2016.11.009

    Figure Lengend Snippet: BPMO-M23D and BPMO-M23D MSTN Administration Induce Substantial Body-wide Dystrophin Restoration (A) Western blot analysis showing dystrophin expression (dys + ) in DIA, GAS, SOL, TA, and heart muscles of BPMO-M23D- and BPMO-M23D MSTN-treated mdx mice. Each lane represents a sample from an individual mouse. Alpha-tubulin (α-tub + ) was used as an internal loading control for western blot. (B) Quantification of dystrophin expression by densitometric analysis of western blot. Following western blot evaluation, the intensity of dys + patterns was scored and normalized to the intensity of corresponding α-tub + patterns, and subsequently it was quantified based on a standard curve of C57 dystrophin. The results were expressed as the percentage of muscle type-matched C57 value (considered as 100%). Data are shown for individual muscle types or as an average of all types. (C and D) Immunostaining detecting dystrophin and laminin expression in treated muscles. Representative images of (C) DIA and (D) TA muscle sections for each group of mice are shown, respectively. Dystrophin-positive fibers were stained in green while laminin-positive fibers were stained in red. Nuclei were stained in blue with DAPI. Scale bars, 100 μm. (E) Quantification of dystrophin intensity levels in DIA, EDL, GAS, SOL, TA, and heart muscles. Following immunostaining for dystrophin, the mean dystrophin intensity was scored by ZEN software and normalized to the mean intensity of laminin detected on the same section. Results were expressed as the percentage of C57 value, considered as 100%. (F and G) Quantification of dystrophin-positive fibers was focused on (F) DIA and (G) TA muscles. The number of dystrophin- and laminin-positive fibers from five random fields of mid-belly muscle sections was counted. Only fibers showing continuous staining of dystrophin along the entire sarcolemma were considered as dystrophin positive and evaluated as the percentage of the number of total fibers (laminin positive) within the same image field. Results were expressed as the percentage of muscle type-matched C57 value, obtained in the same way and considered as 100%. Data in (B) and (E)–(G) are shown as means ± SEM; error bars represent the SEM; n = 10/group. Statistical comparison was two-tailed Student’s t test (*p

    Article Snippet: Primary antibodies were mouse anti-MHC antibodies (1:10, DSHB), including BA-D5 for MHC I, SC-71 for MHC IIA, and BF-F3 for MHC IIB; rabbit anti-laminin antibody (1:300, Abcam) was additionally used.

    Techniques: Western Blot, Expressing, Mouse Assay, Immunostaining, Staining, Software, Two Tailed Test

    BPMO-Mediated Therapy Robustly Improves Hallmarks of Dystrophic Muscles (A and B) Quantification of centrally nucleated fibers in (A) DIA and (B) TA muscles, respectively. Results are expressed as the percentage of the total fibers. (C and D) Frequency distribution of the minimal Feret’s diameter of (C) DIA and (D) TA myofibers. Muscle sections were immunostained for laminin. The minimal Feret’s diameter was semi-automatically measured by ZEN imaging analysis software. Incomplete fibers were excluded from the analysis. The frequency distribution of the Feret’s diameter was analyzed by Prism5. Data are shown as the percentage of the total fiber number. (E and F) Mean of the Feret’s diameter is displayed for (E) DIA and (F) TA fibers, respectively. (G and H) Evaluation of muscle fibrosis in (G) DIA and (H) TA cross sections. Immunostaining for collagen VI was performed. Representative mosaic images showing the entire sections and images at higher magnification are shown. Scale bars, 100 μm (enlarged images of both G and H), 500 μm (G), and 1,000 μm (H). (I and J) Quantification of muscle fibrosis. Following immunostaining for collagen VI, the mean intensity of collagen VI was measured by ZEN software and expressed as the percentage of C57 values (considered as 100%). (K) Immunostaining of DIA sections using antibodies detecting four MHC fiber types. Representative mosaic images of all treatment groups are shown. MHC I fibers were stained in red, MHC IIA fibers were stained in green, MHC IIB fibers were stained in blue, and MHC IIX fibers were unstained. Immunostaining for laminin was used for identifying the sarcolemma of the myofibers. Scale bars, 500 μm. (L–O) Quantification of MHC fibers in DIA transverse sections. Following immunostaining, mosaic images of the whole-muscle sections were generated using ZEN software. The number of MHC-positive fibers was counted separately using ImageJ software, and it was expressed as the percentage of the total number of all fiber types within each muscle section. Data in (A), (B), (E), (F), (I), (J), and (L)–(O) are shown as means ± SEM; error bars represent the SEM; n = 10/group. Statistical comparison was by one-way ANOVA followed by Bonferroni’s post hoc test (*p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Systemic Antisense Therapeutics for Dystrophin and Myostatin Exon Splice Modulation Improve Muscle Pathology of Adult mdx Mice

    doi: 10.1016/j.omtn.2016.11.009

    Figure Lengend Snippet: BPMO-Mediated Therapy Robustly Improves Hallmarks of Dystrophic Muscles (A and B) Quantification of centrally nucleated fibers in (A) DIA and (B) TA muscles, respectively. Results are expressed as the percentage of the total fibers. (C and D) Frequency distribution of the minimal Feret’s diameter of (C) DIA and (D) TA myofibers. Muscle sections were immunostained for laminin. The minimal Feret’s diameter was semi-automatically measured by ZEN imaging analysis software. Incomplete fibers were excluded from the analysis. The frequency distribution of the Feret’s diameter was analyzed by Prism5. Data are shown as the percentage of the total fiber number. (E and F) Mean of the Feret’s diameter is displayed for (E) DIA and (F) TA fibers, respectively. (G and H) Evaluation of muscle fibrosis in (G) DIA and (H) TA cross sections. Immunostaining for collagen VI was performed. Representative mosaic images showing the entire sections and images at higher magnification are shown. Scale bars, 100 μm (enlarged images of both G and H), 500 μm (G), and 1,000 μm (H). (I and J) Quantification of muscle fibrosis. Following immunostaining for collagen VI, the mean intensity of collagen VI was measured by ZEN software and expressed as the percentage of C57 values (considered as 100%). (K) Immunostaining of DIA sections using antibodies detecting four MHC fiber types. Representative mosaic images of all treatment groups are shown. MHC I fibers were stained in red, MHC IIA fibers were stained in green, MHC IIB fibers were stained in blue, and MHC IIX fibers were unstained. Immunostaining for laminin was used for identifying the sarcolemma of the myofibers. Scale bars, 500 μm. (L–O) Quantification of MHC fibers in DIA transverse sections. Following immunostaining, mosaic images of the whole-muscle sections were generated using ZEN software. The number of MHC-positive fibers was counted separately using ImageJ software, and it was expressed as the percentage of the total number of all fiber types within each muscle section. Data in (A), (B), (E), (F), (I), (J), and (L)–(O) are shown as means ± SEM; error bars represent the SEM; n = 10/group. Statistical comparison was by one-way ANOVA followed by Bonferroni’s post hoc test (*p

    Article Snippet: Primary antibodies were mouse anti-MHC antibodies (1:10, DSHB), including BA-D5 for MHC I, SC-71 for MHC IIA, and BF-F3 for MHC IIB; rabbit anti-laminin antibody (1:300, Abcam) was additionally used.

    Techniques: Imaging, Software, Immunostaining, Staining, Generated

    Infiltration of human breast cancer tumors by mouse TAMs . ( a ) BT474-M1 and ( b ) HCC1954 tumors were established in CB17-SCID-beige mice. Tumors were harvested at 4 weeks after tumor cell injection and sections analyzed for vascularization using antibodies against mouse CD31 (endothelial marker); for tumor ECM, using antibodies against mouse laminin; for leukocyte infiltration using antibodies against mouse CD45; and for TAM infiltration using antibodies against mouse F4/80. Bar = 40 µm. TAM, tumor-associated macrophage.

    Journal: Molecular Therapy

    Article Title: Controlled Extracellular Matrix Degradation in Breast Cancer Tumors Improves Therapy by Trastuzumab

    doi: 10.1038/mt.2010.256

    Figure Lengend Snippet: Infiltration of human breast cancer tumors by mouse TAMs . ( a ) BT474-M1 and ( b ) HCC1954 tumors were established in CB17-SCID-beige mice. Tumors were harvested at 4 weeks after tumor cell injection and sections analyzed for vascularization using antibodies against mouse CD31 (endothelial marker); for tumor ECM, using antibodies against mouse laminin; for leukocyte infiltration using antibodies against mouse CD45; and for TAM infiltration using antibodies against mouse F4/80. Bar = 40 µm. TAM, tumor-associated macrophage.

    Article Snippet: The following antibodies were used in immunohistochemical studies: rat anti-mouse CD31 (BD Pharmingen, San Jose, CA), rat anti-mouse CD45 (BD Pharmingen), rat anti-mouse F4/80 (Abcam, Cambridge, MA), goat antihuman collagen IV (Southern Technology, Longwood, FL), rabbit anti-laminin (DAKO, Glostrup, Denmark), mouse anti-Her2/ neu (Abcam), anti-mouse Ig-AlexaFluor 488 (green) (Invitrogen, Molecular Probes, Eugene OR), anti-rabbit Ig-AlexaFluor 568 (red) (Molecular Probes).

    Techniques: Mouse Assay, Injection, Marker

    Immunofluorescence microscopy for nephrin (green) in glomeruli of WT-C (a and e), WT-R (b and f), TG-C (c and g), and TG-R rats (d and h) at 12 weeks of age. Double-labeled immunostaining for laminin (red) was carried out to locate the glomerular capillary wall. Experimental groups were as follows: wild-type littermates (WT) without risedronate administration (WT-C), WT rats with risedronate administration (WT-R), Pit-1 TG rats with risedronate administration (TG-R), and Pit-1 TG rats without risedronate administration (TG-C). Bar = 50 μ m (a, b, c, and d). Bar = 25 μ m (e, f, g, and h).

    Journal: International Journal of Endocrinology

    Article Title: Risedronate Attenuates Podocyte Injury in Phosphate Transporter-Overexpressing Rats

    doi: 10.1155/2019/4194853

    Figure Lengend Snippet: Immunofluorescence microscopy for nephrin (green) in glomeruli of WT-C (a and e), WT-R (b and f), TG-C (c and g), and TG-R rats (d and h) at 12 weeks of age. Double-labeled immunostaining for laminin (red) was carried out to locate the glomerular capillary wall. Experimental groups were as follows: wild-type littermates (WT) without risedronate administration (WT-C), WT rats with risedronate administration (WT-R), Pit-1 TG rats with risedronate administration (TG-R), and Pit-1 TG rats without risedronate administration (TG-C). Bar = 50 μ m (a, b, c, and d). Bar = 25 μ m (e, f, g, and h).

    Article Snippet: The primary antibodies used were as follows: monoclonal anti-nephrin antibody (5-1-6; courtesy of Dr. H. Kawachi, Niigata University, Niigta, Japan) [ ]; murine monoclonal anti-desmin antibody (clone D-33; Dako Cytomation, Glostrup, Denmark); rabbit anti-connexin43 antibody (Sigma-Aldrich, St Louis, MO, USA); murine monoclonal anti-ZO-1 antibody (Zymed Laboratories, South San Francisco, CA, USA); and rabbit anti-laminin antibody (Dako Cytomation).

    Techniques: Immunofluorescence, Microscopy, Labeling, Immunostaining

    Immunofluorescence microscopy for desmin (green) in glomeruli of WT-C (a and e), WT-R (b and f), TG-C (c and g), and TG-R rats (d and h) at 12 weeks of age. Double-labeled immunostaining for laminin (red) was carried out to locate the glomerular capillary wall. Experimental groups were as follows: wild-type littermates (WT) without risedronate administration (WT-C), WT rats with risedronate administration (WT-R), Pit-1 TG rats with risedronate administration (TG-R), and Pit-1 TG rats without risedronate administration (TG-C). Bar = 50 μ m (a, b, c, and d). Bar = 25 μ m (e, f, g, and h).

    Journal: International Journal of Endocrinology

    Article Title: Risedronate Attenuates Podocyte Injury in Phosphate Transporter-Overexpressing Rats

    doi: 10.1155/2019/4194853

    Figure Lengend Snippet: Immunofluorescence microscopy for desmin (green) in glomeruli of WT-C (a and e), WT-R (b and f), TG-C (c and g), and TG-R rats (d and h) at 12 weeks of age. Double-labeled immunostaining for laminin (red) was carried out to locate the glomerular capillary wall. Experimental groups were as follows: wild-type littermates (WT) without risedronate administration (WT-C), WT rats with risedronate administration (WT-R), Pit-1 TG rats with risedronate administration (TG-R), and Pit-1 TG rats without risedronate administration (TG-C). Bar = 50 μ m (a, b, c, and d). Bar = 25 μ m (e, f, g, and h).

    Article Snippet: The primary antibodies used were as follows: monoclonal anti-nephrin antibody (5-1-6; courtesy of Dr. H. Kawachi, Niigata University, Niigta, Japan) [ ]; murine monoclonal anti-desmin antibody (clone D-33; Dako Cytomation, Glostrup, Denmark); rabbit anti-connexin43 antibody (Sigma-Aldrich, St Louis, MO, USA); murine monoclonal anti-ZO-1 antibody (Zymed Laboratories, South San Francisco, CA, USA); and rabbit anti-laminin antibody (Dako Cytomation).

    Techniques: Immunofluorescence, Microscopy, Labeling, Immunostaining

    Engineered microvasculature exhibits appropriate endothelial barrier function a) Schematic showing while seeded endothelial cells initially adhered to gelatin, they appropriately assembly basement membrane during culture. b) Representative 3D confocal microscopy immunostaining images of the adherens junction protein VE-cadherin and the basement membrane proteins laminin and collagen-IV, self-deposited by HUVECs after 14 days of culture. c) A representative stitched composite of epi-fluorescence images after 15-minute perfusion of BSA-AF594. d) Fluorescence intensity of the BSA-AF594 along the linescan across the engineered microvasculature in panel b. e) Higher magnification view of the engineered microvasculature demonstrating that the system is impermeable to BSA under physiologic flow conditions. f) In acellular (non-endothelialized) microsystems, significant diffusion of BSA occured as early as 5 minutes after perfusion (dashed lines define the microchannel borders). g) Representative plots of BSA-AF594 fluorescence intensity over time in permeability assays for both acellular and endothelialized microdevices. h) Quantified apparent permeability (P a ) of the engineered microvasculature to BSA remained similar across different endothelial cell types, and is approximately 40× less than that in acellular microchannels. Data was plotted as the mean ± s.d. with n=4 independent biological replicates expect for HLMVECs (n=3). P -values were calculated using one-way ANOVA with Bonferroni’s post hoc test (*** P

    Journal: Nature biomedical engineering

    Article Title: Microvasculature-on-a-chip for the long-term study of endothelial barrier dysfunction and microvascular obstruction in disease

    doi: 10.1038/s41551-018-0224-z

    Figure Lengend Snippet: Engineered microvasculature exhibits appropriate endothelial barrier function a) Schematic showing while seeded endothelial cells initially adhered to gelatin, they appropriately assembly basement membrane during culture. b) Representative 3D confocal microscopy immunostaining images of the adherens junction protein VE-cadherin and the basement membrane proteins laminin and collagen-IV, self-deposited by HUVECs after 14 days of culture. c) A representative stitched composite of epi-fluorescence images after 15-minute perfusion of BSA-AF594. d) Fluorescence intensity of the BSA-AF594 along the linescan across the engineered microvasculature in panel b. e) Higher magnification view of the engineered microvasculature demonstrating that the system is impermeable to BSA under physiologic flow conditions. f) In acellular (non-endothelialized) microsystems, significant diffusion of BSA occured as early as 5 minutes after perfusion (dashed lines define the microchannel borders). g) Representative plots of BSA-AF594 fluorescence intensity over time in permeability assays for both acellular and endothelialized microdevices. h) Quantified apparent permeability (P a ) of the engineered microvasculature to BSA remained similar across different endothelial cell types, and is approximately 40× less than that in acellular microchannels. Data was plotted as the mean ± s.d. with n=4 independent biological replicates expect for HLMVECs (n=3). P -values were calculated using one-way ANOVA with Bonferroni’s post hoc test (*** P

    Article Snippet: The fixed engineered microvasculature was then blocked by perfusion of 1% BSA solution, followed by perfusion of the primary antibody, mouse anti-human VE-cadherin (1:100, F8, Santa Cruz Biotech), mouse anti-human collagen IV (1:100, abcam), or rabbit anti-human laminin (1:100, abcam).

    Techniques: Confocal Microscopy, Immunostaining, Fluorescence, Flow Cytometry, Diffusion-based Assay, Permeability