monoclonal mouse anti human smooth muscle actin  (Agilent technologies)

Journal: Scientific Reports

Article Title: Antifibrotic effect of disulfiram on bleomycin-induced lung fibrosis in mice and its impact on macrophage infiltration

doi: 10.1038/s41598-024-71770-z

Figure Lengend Snippet: Immunohistochemical analysis of BLM-induced lung fibrosis in mice. ( A – F ) Representative immunostaining images for type I collagen ( A – C ), α-SMA ( D – F ), and F4/80 ( G – I ) in the lung tissue of untreated ( A , D , G ), DSF non-treated ( B , E , H ) and DSF-treated ( C , F , I ) mice on day 28. Higher-magnification images in the lower left correspond to the areas indicated by blue squares. Scale bars = 500 μm. ( J , K ) Percentage of type I collagen positive and α-SMA positive area in lung tissue on day 28. ( L ) Number of F4/80 + cells in the lung tissue on day 28 counted in immunostaining images ( G – I ). n = 5 per group. * P < 0.05, ** P < 0.01 (Tukey’s test). ns, not significant.

Article Snippet: We used the following primary antibodies: anti-human α-smooth muscle actin (α-SMA) mouse monoclonal antibody (1:250 dilution, Dako, Glostrup, Denmark), anti-human CD34 mouse monoclonal antibody (1:100 dilution, Nichirei Bioscience, Tokyo, Japan), anti-human CD68 mouse monoclonal antibody (1:200 dilution, Dako, Glostrup, Denmark), anti-human/mouse S100A4 rabbit monoclonal antibody (1:100 dilution, Abcam, Cambridge, UK), and anti-human FROUNT goat polyclonal antibody (1:150 dilution, Everest biotech, Oxfordshire, UK), followed by peroxidase-labelled secondary antibodies, and incubated with Opal fluorophore Opal 690, Opal 540, Opal 570, Opal 620, Opal 520, respectively.

Techniques: Immunohistochemical staining, Immunostaining

Journal: Scientific Reports

Article Title: Antifibrotic effect of disulfiram on bleomycin-induced lung fibrosis in mice and its impact on macrophage infiltration

doi: 10.1038/s41598-024-71770-z

Figure Lengend Snippet: Analysis of multi-fluorescent staining of lung tissues of IPF. ( A , B ) Representative immunofluorescent staining images in normal lung ( A ) and in fibrotic areas of the lungs of IPF patients ( B ). DAPI is shown in pink, α-SMA in brown, CD34 in light blue, CD68 in green, S100A4 in red, and FROUNT in blue. Higher-magnification images on the right correspond to the area indicated with dashed squares. White arrows indicate CD68 + macrophages in normal lung. White arrowheads indicate S100A4 + CD68 + macrophages and yellow arrowheads indicate S100A4 - CD68 + macrophages in fibrotic areas of the lungs of IPF patients. Scale bars = 25 μm. ( C ) S100A4 signals of each CD68 + macrophage in normal lung tissues and in fibrotic areas of the lungs of IPF patients. ( D ) FROUNT signals in S100A4 - CD68 + macrophages and S100A4 + CD68 + macrophages in fibrotic areas of the lungs of IPF patients. n = 5 or 6 per group. ** P < 0.01 (unpaired t-test).

Article Snippet: We used the following primary antibodies: anti-human α-smooth muscle actin (α-SMA) mouse monoclonal antibody (1:250 dilution, Dako, Glostrup, Denmark), anti-human CD34 mouse monoclonal antibody (1:100 dilution, Nichirei Bioscience, Tokyo, Japan), anti-human CD68 mouse monoclonal antibody (1:200 dilution, Dako, Glostrup, Denmark), anti-human/mouse S100A4 rabbit monoclonal antibody (1:100 dilution, Abcam, Cambridge, UK), and anti-human FROUNT goat polyclonal antibody (1:150 dilution, Everest biotech, Oxfordshire, UK), followed by peroxidase-labelled secondary antibodies, and incubated with Opal fluorophore Opal 690, Opal 540, Opal 570, Opal 620, Opal 520, respectively.

Techniques: Staining

Journal: Scientific Reports

Article Title: Antifibrotic effect of disulfiram on bleomycin-induced lung fibrosis in mice and its impact on macrophage infiltration

doi: 10.1038/s41598-024-71770-z

Figure Lengend Snippet: Immunohistochemical analysis of BLM-induced lung fibrosis in mice. ( A – F ) Representative immunostaining images for type I collagen ( A – C ), α-SMA ( D – F ), and F4/80 ( G – I ) in the lung tissue of untreated ( A , D , G ), DSF non-treated ( B , E , H ) and DSF-treated ( C , F , I ) mice on day 28. Higher-magnification images in the lower left correspond to the areas indicated by blue squares. Scale bars = 500 μm. ( J , K ) Percentage of type I collagen positive and α-SMA positive area in lung tissue on day 28. ( L ) Number of F4/80 + cells in the lung tissue on day 28 counted in immunostaining images ( G – I ). n = 5 per group. * P < 0.05, ** P < 0.01 (Tukey’s test). ns, not significant.

Article Snippet: We used the following primary antibodies: anti-human α-smooth muscle actin (α-SMA) mouse monoclonal antibody (1:250 dilution, Dako, Glostrup, Denmark), anti-human CD34 mouse monoclonal antibody (1:100 dilution, Nichirei Bioscience, Tokyo, Japan), anti-human CD68 mouse monoclonal antibody (1:200 dilution, Dako, Glostrup, Denmark), anti-human/mouse S100A4 rabbit monoclonal antibody (1:100 dilution, Abcam, Cambridge, UK), and anti-human FROUNT goat polyclonal antibody (1:150 dilution, Everest biotech, Oxfordshire, UK), followed by peroxidase-labelled secondary antibodies, and incubated with Opal fluorophore Opal 690, Opal 540, Opal 570, Opal 620, Opal 520, respectively.

Techniques: Immunohistochemical staining, Immunostaining

Journal: Scientific Reports

Article Title: Antifibrotic effect of disulfiram on bleomycin-induced lung fibrosis in mice and its impact on macrophage infiltration

doi: 10.1038/s41598-024-71770-z

Figure Lengend Snippet: Analysis of multi-fluorescent staining of lung tissues of IPF. ( A , B ) Representative immunofluorescent staining images in normal lung ( A ) and in fibrotic areas of the lungs of IPF patients ( B ). DAPI is shown in pink, α-SMA in brown, CD34 in light blue, CD68 in green, S100A4 in red, and FROUNT in blue. Higher-magnification images on the right correspond to the area indicated with dashed squares. White arrows indicate CD68 + macrophages in normal lung. White arrowheads indicate S100A4 + CD68 + macrophages and yellow arrowheads indicate S100A4 - CD68 + macrophages in fibrotic areas of the lungs of IPF patients. Scale bars = 25 μm. ( C ) S100A4 signals of each CD68 + macrophage in normal lung tissues and in fibrotic areas of the lungs of IPF patients. ( D ) FROUNT signals in S100A4 - CD68 + macrophages and S100A4 + CD68 + macrophages in fibrotic areas of the lungs of IPF patients. n = 5 or 6 per group. ** P < 0.01 (unpaired t-test).

Article Snippet: We used the following primary antibodies: anti-human α-smooth muscle actin (α-SMA) mouse monoclonal antibody (1:250 dilution, Dako, Glostrup, Denmark), anti-human CD34 mouse monoclonal antibody (1:100 dilution, Nichirei Bioscience, Tokyo, Japan), anti-human CD68 mouse monoclonal antibody (1:200 dilution, Dako, Glostrup, Denmark), anti-human/mouse S100A4 rabbit monoclonal antibody (1:100 dilution, Abcam, Cambridge, UK), and anti-human FROUNT goat polyclonal antibody (1:150 dilution, Everest biotech, Oxfordshire, UK), followed by peroxidase-labelled secondary antibodies, and incubated with Opal fluorophore Opal 690, Opal 540, Opal 570, Opal 620, Opal 520, respectively.

Techniques: Staining

Journal: Scientific Reports

Article Title: Antifibrotic effect of disulfiram on bleomycin-induced lung fibrosis in mice and its impact on macrophage infiltration

doi: 10.1038/s41598-024-71770-z

Figure Lengend Snippet: Immunohistochemical analysis of BLM-induced lung fibrosis in mice. ( A – F ) Representative immunostaining images for type I collagen ( A – C ), α-SMA ( D – F ), and F4/80 ( G – I ) in the lung tissue of untreated ( A , D , G ), DSF non-treated ( B , E , H ) and DSF-treated ( C , F , I ) mice on day 28. Higher-magnification images in the lower left correspond to the areas indicated by blue squares. Scale bars = 500 μm. ( J , K ) Percentage of type I collagen positive and α-SMA positive area in lung tissue on day 28. ( L ) Number of F4/80 + cells in the lung tissue on day 28 counted in immunostaining images ( G – I ). n = 5 per group. * P < 0.05, ** P < 0.01 (Tukey’s test). ns, not significant.

Article Snippet: We used the following primary antibodies: anti-human α-smooth muscle actin (α-SMA) mouse monoclonal antibody (1:250 dilution, Dako, Glostrup, Denmark), anti-human CD34 mouse monoclonal antibody (1:100 dilution, Nichirei Bioscience, Tokyo, Japan), anti-human CD68 mouse monoclonal antibody (1:200 dilution, Dako, Glostrup, Denmark), anti-human/mouse S100A4 rabbit monoclonal antibody (1:100 dilution, Abcam, Cambridge, UK), and anti-human FROUNT goat polyclonal antibody (1:150 dilution, Everest biotech, Oxfordshire, UK), followed by peroxidase-labelled secondary antibodies, and incubated with Opal fluorophore Opal 690, Opal 540, Opal 570, Opal 620, Opal 520, respectively.

Techniques: Immunohistochemical staining, Immunostaining

Journal: Scientific Reports

Article Title: Antifibrotic effect of disulfiram on bleomycin-induced lung fibrosis in mice and its impact on macrophage infiltration

doi: 10.1038/s41598-024-71770-z

Figure Lengend Snippet: Analysis of multi-fluorescent staining of lung tissues of IPF. ( A , B ) Representative immunofluorescent staining images in normal lung ( A ) and in fibrotic areas of the lungs of IPF patients ( B ). DAPI is shown in pink, α-SMA in brown, CD34 in light blue, CD68 in green, S100A4 in red, and FROUNT in blue. Higher-magnification images on the right correspond to the area indicated with dashed squares. White arrows indicate CD68 + macrophages in normal lung. White arrowheads indicate S100A4 + CD68 + macrophages and yellow arrowheads indicate S100A4 - CD68 + macrophages in fibrotic areas of the lungs of IPF patients. Scale bars = 25 μm. ( C ) S100A4 signals of each CD68 + macrophage in normal lung tissues and in fibrotic areas of the lungs of IPF patients. ( D ) FROUNT signals in S100A4 - CD68 + macrophages and S100A4 + CD68 + macrophages in fibrotic areas of the lungs of IPF patients. n = 5 or 6 per group. ** P < 0.01 (unpaired t-test).

Article Snippet: We used the following primary antibodies: anti-human α-smooth muscle actin (α-SMA) mouse monoclonal antibody (1:250 dilution, Dako, Glostrup, Denmark), anti-human CD34 mouse monoclonal antibody (1:100 dilution, Nichirei Bioscience, Tokyo, Japan), anti-human CD68 mouse monoclonal antibody (1:200 dilution, Dako, Glostrup, Denmark), anti-human/mouse S100A4 rabbit monoclonal antibody (1:100 dilution, Abcam, Cambridge, UK), and anti-human FROUNT goat polyclonal antibody (1:150 dilution, Everest biotech, Oxfordshire, UK), followed by peroxidase-labelled secondary antibodies, and incubated with Opal fluorophore Opal 690, Opal 540, Opal 570, Opal 620, Opal 520, respectively.

Techniques: Staining