mouse anti mf20  (Developmental Studies Hybridoma Bank)

Journal: Frontiers in Immunology

Article Title: A biomimetic model composed of injectable 3D muscle-like tissue, stromal and immune cells for recapitulating the rapid immune signature predictive of mRNA vaccine immunogenicity

doi: 10.3389/fimmu.2025.1651095

Figure Lengend Snippet: Expression of SARS-CoV-2 spike (S) protein in fibroblasts and muscle models in vitro stimulated with BNT162b2 vaccine. Fibroblasts (A) , myogenic progenitors (MP) (B) , myotubes (MT) (C) and 3D muscle-like tissue (3D-MT) (D) were left not stimulated (NS) or treated with BNT162b2 vaccine (VAX, 1 µg/mL) for 24 h (H) . Immunofluorescence analysis was conducted by confocal microscopy to assess the expression of S protein. Fibroblasts were stained with DAPI (blue) to identify nuclei and with an anti-S antibody (Ab) (green) (A) . For MP staining, in addition to DAPI (blue) and anti-S Ab (green), an anti-phalloidin Ab (red) was used as a marker of undifferentiated muscle cells (B) , while for MT and 3D-MT, staining with anti-myosin heavy chain (MyHC) Ab (red) was employed as a differentiation marker. Representative images, out of three experiments ( n = 3) separately performed, are shown in panels (A–D) . Scale bar: 20 μm.

Article Snippet: Afterward, 2D MP were incubated for 1 h at RT with Phalloidin Alexa Fluor 555 (Invitrogen, Thermo Fisher Scientific, A12379) for labeling F-actin of cytoskeleton, while MT and 3D-MT were incubated for 1 h and o.n. at RT, respectively, with anti-myosin heavy chain (anti-MF20) mouse primary antibody (DSHB, AB_2147781) diluted 1:200 to assess terminal myogenic differentiation.

Techniques: Expressing, In Vitro, Immunofluorescence, Confocal Microscopy, Staining, Marker

Journal: Cells

Article Title: Dynamic Expression and Functional Implications of the Cell Polarity Gene, Dchs1, During Cardiac Development

doi: 10.3390/cells14110774

Figure Lengend Snippet: DCHS1-HA expression at embryonic day 11.5. ( A ) IHC of E11.5 embryos for HA (red) and CD31 (green) showing co-localization of Dchs1-HA and a subset of endothelial cells and expression within the pericardium. Scale bar = 170 μm ( B ) IHC of HA (red) and MF20 (green) showing no detectable expression of DCHS1 in cardiomyocytes. Scale bar = 170 μm ( C ) IHC of HA (red) and MF20 (green) in the atrioventricular (AV) cushions showed robust DCHS1 expression as well as expression within the epicardium. Scale bar = 80 μm ( D ) Western analyses of heart lysates at E11.5 probed for HA showing full-length (>250 kDa) and smaller fragments. ( E ) Ponceau-S staining of western blot showing equal loading of proteins. Lumen (Lum), myocardium (Myo), epicardium (Epi), pericardium (Peri), atrioventricular cushions (AVC).

Article Snippet: The following primary antibodies and reagents were used for immunostaining: rabbit anti-HA antibody (Cell Signaling, # 3724; Danvers, MN, USA) at a dilution of 1:250, rat anti-CD31 antibody (Dianova, # DIA-310; Hamburg, Germany) at 1:100, and mouse anti-MF20 antibody (Developmental Studies Hybridoma Bank, Concentrate 0.1 mL; Iowa City, IA, USA) at 1:500.

Techniques: Expressing, Western Blot, Staining

Journal: Cells

Article Title: Dynamic Expression and Functional Implications of the Cell Polarity Gene, Dchs1, During Cardiac Development

doi: 10.3390/cells14110774

Figure Lengend Snippet: DCHS1-HA expression at embryonic day 13.5. ( A ) Left ventricular free wall cardiac tissue was co-stained with HA (red) and CD31 (green), showing significant overlap (yellow), demonstrating co-expression within endothelial cells. ( B ) No overlap in staining was detected between MF20 (green) and DCHS1 (red). ( C ) Atrioventricular (AV) cushions demonstrated a gradient of DCHS1 expression, with the highest levels within the subendocardium (**) and lower levels closer to the AV myocardium (*). ( D ) Interventricular septum showed relatively low levels of endocardial DCHS1 expression at this time point. ( E ) Western analyses of heart lysates at E13.5 probed for HA showing full-length (>250 kDa) and smaller fragments. ( F ) Ponceau-S staining of western blot showing equal loading of proteins. Lumen (Lum), trabeculated myocardium (T), myocardium (Myo), epicardium (Epi), and atrioventricular cushions (AVC).

Article Snippet: The following primary antibodies and reagents were used for immunostaining: rabbit anti-HA antibody (Cell Signaling, # 3724; Danvers, MN, USA) at a dilution of 1:250, rat anti-CD31 antibody (Dianova, # DIA-310; Hamburg, Germany) at 1:100, and mouse anti-MF20 antibody (Developmental Studies Hybridoma Bank, Concentrate 0.1 mL; Iowa City, IA, USA) at 1:500.

Techniques: Expressing, Staining, Western Blot

Journal: Cells

Article Title: Dynamic Expression and Functional Implications of the Cell Polarity Gene, Dchs1, During Cardiac Development

doi: 10.3390/cells14110774

Figure Lengend Snippet: DCHS1-HA expression during fetal gestation. ( A ) Left ventricular free wall cardiac tissue was co-stained with HA (red) and CD31 (green), demonstrating significant overlap. At this timepoint, some non-endocardial, non-MF20 (green) staining is evident (arrows). Epicardial expression of DCHS1-HA is prominent (arrowheads). ( B ) HA (red) and MF20 (green) in the LV free wall showing no detectable overlap in expression. ( C ) IHC for HA (red) and MF20 (green) showing a gradient expression of DCHS1-HA, with the highest levels within the mitral valve subendocardium (**) and lower levels closer to the myocardium (*). ( D ) tissue from the interventricular septum was co-stained with HA (for DCHS1-HA) in red and CD31 (endothelial cell marker) in green. ( E ) Whole-heart tissue lysates of pups from an E18.5 litter were probed blindly for HA (to visualize DCHS1-HA), with observed bands highlighted with arrowheads. Arrow represents non-specific (n.s.) antibody staining on the blot. ( F ) Ponceau-S total protein staining of western blot analyzed in ( E ). Labeled abbreviations: lumen (Lum), right ventricular lumen (RV Lum), trabeculated myocardium (T), myocardium (Myo), epicardium (Epi), and pericardium (Peri).

Article Snippet: The following primary antibodies and reagents were used for immunostaining: rabbit anti-HA antibody (Cell Signaling, # 3724; Danvers, MN, USA) at a dilution of 1:250, rat anti-CD31 antibody (Dianova, # DIA-310; Hamburg, Germany) at 1:100, and mouse anti-MF20 antibody (Developmental Studies Hybridoma Bank, Concentrate 0.1 mL; Iowa City, IA, USA) at 1:500.

Techniques: Expressing, Staining, Marker, Western Blot, Labeling

Journal: Cells

Article Title: Dynamic Expression and Functional Implications of the Cell Polarity Gene, Dchs1, During Cardiac Development

doi: 10.3390/cells14110774

Figure Lengend Snippet: Dchs1 Expression at post-natal day 0 (P0). ( A ) IHC of left ventricular free wall was performed for HA (red) and CD31 (green) or ( B ) MF20 (green). Consistent with earlier timepoints, DCHS1 was not observed on cardiomyocytes but was detected on endothelial cells. Robust and extensive staining was also observed throughout the myocardium in cells that are negative for CD31 and MF20 (indicated by arrows in 1A), likely representing a fibroblast population. ( C ) IHC in mitral valve displayed extensive staining throughout the leaflet, with the most abundant expression within the subendocardial mesenchyme. ( D ) IHC in the interventricular septum was similar with the LV free wall in that DCHS1 is co-expressed with CD31 as well as prevalent in a non-myocyte, non-endothelial cell population. ( E ) Western analyses of whole-heart lysates revealed maintenance of the full-length protein and presence of a small-molecular-weight fragment (arrowheads). Arrow represents non-specific (n.s.) antibody staining on the blot. ( F ) Ponceau-S total protein staining demonstrates equal loading between samples. Lumen (Lum), right/left ventricular lumen (RV/LV, respectively, lumen), trabeculated myocardium (T), myocardium (Myo), epicardium (Epi), mitral valve (MV).

Article Snippet: The following primary antibodies and reagents were used for immunostaining: rabbit anti-HA antibody (Cell Signaling, # 3724; Danvers, MN, USA) at a dilution of 1:250, rat anti-CD31 antibody (Dianova, # DIA-310; Hamburg, Germany) at 1:100, and mouse anti-MF20 antibody (Developmental Studies Hybridoma Bank, Concentrate 0.1 mL; Iowa City, IA, USA) at 1:500.

Techniques: Expressing, Staining, Western Blot, Molecular Weight

Journal: Cells

Article Title: Dynamic Expression and Functional Implications of the Cell Polarity Gene, Dchs1, During Cardiac Development

doi: 10.3390/cells14110774

Figure Lengend Snippet: Adolescent and adult Dchs1 expression. ( A , B ) Post-natal LV free wall and mitral valves at day 7 (P7) and P21 mice were stained with HA (red) and MF20 (green). ( C , D ) Western analyses from whole-heart tissue lysates of pups from P7 and P21 probed for HA. Full length (>250 kDa) was barely detectable at this stage, and reduction in the smaller molecular weight is also apparent at both P7 and P21 (arrowheads). At these timepoints, additional smaller fragments are observed, indicating potential degradation of DCHS1. Arrow represents non-specific (n.s.) immunoreactive bands. ( E , F ) Ponceau-S stain showing equivalent protein loading, respectively. Labeled abbreviations: lumen (Lum), trabeculated myocardium (T), myocardium (Myo), epicardium (Epi), mitral valve (MV), and interventricular septum (IVS).

Article Snippet: The following primary antibodies and reagents were used for immunostaining: rabbit anti-HA antibody (Cell Signaling, # 3724; Danvers, MN, USA) at a dilution of 1:250, rat anti-CD31 antibody (Dianova, # DIA-310; Hamburg, Germany) at 1:100, and mouse anti-MF20 antibody (Developmental Studies Hybridoma Bank, Concentrate 0.1 mL; Iowa City, IA, USA) at 1:500.

Techniques: Expressing, Staining, Western Blot, Molecular Weight, Labeling