Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: a Representative images of sections stained with α-smooth muscle actin (α-SMA) and cleaved caspase-3 by immunohistochemistry (IHC) staining from adult rat wounds at day 14 post-injury. Yellow lines outline the scar area. Dashed boxes in the lower magnification images represent the region of interest shown in the higher magnification images below. Scale bars, 100 μm. b Quantification of α-SMA + myofibroblast density in adult rat wounds from ( a ). N = 12 (control) or 14 (FMOD)-treated rats, respectively. c Quantification of cleaved caspase-3 + cell density in adult rat wounds from ( a ). N = 12 rats. d Representative images of sections stained with hematoxylin & eosin (H&E) (first panel), picrosirius red (PSR) coupled with polarized light microscopy (PLM) (second panel), IHC for α-SMA (third panel) and cleaved caspase-3 (fourth panel) from excessive-mechanical-loading wounds of female adult red Duroc pigs at week 8 post-injury. Dermal scar areas are outlined in blue (for the H&E-stained image) or yellow (for PSR-PLM images), while white dotted lines in the PSR-PLM images outline the epidermal edge. The solid green-boxed regions (second panel) represent the region of α-SMA-stained images, while the dotted green-boxed regions (second panel) represent the region of cleaved caspase-3-stained images. Scale bars, 50 (black) or 500 μm (yellow). e Quantification of α-SMA + myofibroblast density in control- vs . FMOD-treated adult pigs from d . f Quantification of cleaved caspase-3 + cell density in control- vs . FMOD-treated adult pigs from ( d ). N = 9 wounds from 3 pigs ( e , f ). All treatments were administrated at the time of surgery. The number of IHC-positively stained cells and nuclei across the entire wound area was counted under a microscope from two centrally bisected sections of each wound sample. The ratio of IHC-positively stained cells to the total number of cells (indicated by the number of nuclei) was then calculated to quantify the density of α−SMA + ( b , e ) or cleaved caspase-3 + ( c and f ) cells. Data presented as mean ± standard deviation (s.d.) overlaying all the data points. P- values were determined by two-tailed unpaired t -tests ( b – f ). * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ).

Techniques: Staining, Immunohistochemistry, Control, Light Microscopy, Microscopy, Standard Deviation, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of human BJ fibroblasts stained with α-smooth muscle actin (α-SMA) by immunofluorescence staining before and after 5.0 ng/mL transforming growth factor (TGF)β1 treatment for 6 days. N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ). N = 3 biological replicates. d Expression of ACTA2 (the gene encoding α-SMA) in BJ fibroblasts before and after fibroblast-myofibroblast conversion. Data were normalized to the ACTA2 level before TGFβ1-induced fibroblast-myofibroblast conversion. N = 3 biological replicates. e Representative images of BJ fibroblast-converted myofibroblasts (BJ-myofibroblasts) with α-SMA and TUNEL staining, accompanied by the respective flow cytometry plots stained with Annexin V-FITC and PI staining. N = 3 (for α-SMA and TUNEL staining) or 4 (for flow cytometry) biological replicates, respectively. f Quantification of apoptotic BJ-myofibroblasts by flow cytometry from ( e ). g Quantification of pyroptotic BJ-myofibroblasts by flow cytometry from ( e ). N = 4 biological replicates ( f , g ). Scale bars, 25 μm. Data presented as mean ± s.d. overlaying all the data points. P- values were determined by two-tailed unpaired t -tests ( c – g ). N.S ., not significant, P > 0.05; * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ).

Techniques: Staining, Immunofluorescence, Flow Cytometry, Expressing, TUNEL Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of the primary NHDF FB-AA36 (derived from a 36-year-old female African–American donor; Supplementary Table ) stained with α-smooth muscle actin (α-SMA) by immunofluorescence staining before and after transforming growth factor (TGF)β1-induced fibroblast-myofibroblast conversion. b Expression of ACTA2 in FB-AA36 fibroblasts before and after fibroblast-myofibroblast conversion. Data were normalized to the ACTA2 level before TGFβ1-induced fibroblast-myofibroblast conversion. c Representative images of NHDF FB-AA36 with α-SMA and TUNEL staining, accompanied by the respective flow cytometry plots stained with Annexin V-FITC and PI staining. d Quantification of NHDF FB-AA36 apoptosis from flow cytometry in ( c ). e Representative images of FB-AA36 fibroblast-converted myofibroblasts with α-SMA and TUNEL staining, accompanied by the respective flow cytometry plots stained with Annexin V-FITC and PI staining. f Quantification of FB-AA36 fibroblast-converted myofibroblast apoptosis from flow cytometry in ( e ). Scale bars, 25 μm. Data presented as mean ± s.d. overlaying all the data points. N = 3 biological replicates; P- values were determined by two-tailed unpaired t -tests ( b – f ). N.S ., not significant, P > 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ).

Techniques: Derivative Assay, Staining, Immunofluorescence, Expressing, TUNEL Assay, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of the human keloid-derived fibroblast KB-AA35 (derived from a 35-year-old female African–American donor; Supplementary Table ) stained with α-smooth muscle actin (α-SMA) by immunofluorescence staining. Scale bar, 25 μm. b Expression of ACTA2 in FB-AA36 fibroblasts and KB-AA35 fibroblasts. Data were normalized to the ACTA2 level of FB-AA36 fibroblasts. c Quantification of KB-AA35 fibroblast apoptosis by flow cytometry with Annexin V-FITC and PI staining. d Expression of IL1B in KB-AA35 fibroblasts with or without FMOD treatment. Data were normalized to the IL1B level without FMOD treatment. e Active IL1β production in KB-AA35 fibroblasts with or without FMOD treatment. Data presented as mean ± s.d. overlaying all the data points. N = 3 biological replicates; P- values were determined by two-tailed unpaired t -tests ( b – e ). * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ).

Techniques: Derivative Assay, Staining, Immunofluorescence, Expressing, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of normal skin (from abdomen), keloid (from abdomen), and hypertrophic scar (from left shoulder) tissues collected from the donor with the National Disease Research Interchange (NDRI) reference ID 1914138 with α-smooth muscle actin (α-SMA) staining by immunohistochemistry (IHC; upper), and the representative images of dermal fibroblasts isolated from these tissues with α-SMA staining by immunofluorescence staining (lower). b Expression of ACTA2 in dermal fibroblasts derived from tissues described from ( a ). Data were normalized to the ACTA2 level of fibroblasts isolated from the normal skin. c Representative flow cytometry plots of fibroblasts derived from tissues described in ( a ) with Annexin V-FITC and PI staining. d Quantification of apoptotic dermal fibroblasts by flow cytometry from ( c ). e The influence of FMOD on cell apoptosis with and without IL1β was calculated from ( d ). Scale bars, 50 μm (black) or 25 μm (white). Data presented as mean ± s.d. overlaying all the data points. N = 3 biological replicates; P -values were determined by two-tailed unpaired t -tests ( b – e ). N.S ., not significant, P > 0.05; * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ).

Techniques: Staining, Immunohistochemistry, Isolation, Immunofluorescence, Expressing, Derivative Assay, Flow Cytometry, Two Tailed Test