Journal: bioRxiv

Article Title: Monocytes Strongly Induce (MYO)Fibroblast Contraction in a New 3D Skin Model to Understand the Inflammation-Fibrosis Axis in Systemic Sclerosis

doi: 10.64898/2026.02.12.705496

Figure Lengend Snippet: ( A ) Sorted CD45-cells (fibroblasts) were analyzed for expression of myofibroblast-related genes after co-culture with monocytes in collagen hydrogels. Values (mean ± SEM) indicate relative gene expression (−ΔCt) measured with qPCR. Reference genes used were GAPDH and RPS27A. Analyses were done by two-way ANOVA with Sidak’s multiple comparisons test (**p<0.01, ***p<0.001, ****p<0.0001). ( B ) Representative immunohistochemistry staining showing robust expression of α-SMA (alpha-smooth muscle actin) (left) and fibroblast activation protein (FAP) (right) in the hydrogels containing only fibroblasts (top) compared to those co-culture with monocytes (bottom). The small blue nuclei in the bottom panels are monocytes. (Myo)fibroblasts are clearly positive for both markers (black arrows), with a typically elongated shape.

Article Snippet: Sections were then incubated with primary rabbit –anti-human alpha-smooth muscle actin (α-SMA) (1:200) (ab5694, Abcam, UK), – fibroblast activation protein (FAP) (1:100) (ab207178, Abcam), – Phospho-SMAD2 (1:100) (#3108L, CellSignaling, USA), – Phospho-Stat3 (1:50) (#9145, CellSignaling), or mouse anti-human – CD68 (#MCA1815, BioRad, USA) (1:200) antibodies, for 1 hour at room temperature.

Techniques: Expressing, Co-Culture Assay, Gene Expression, Immunohistochemistry, Staining, Activation Assay

Journal: bioRxiv

Article Title: Monocytes Strongly Induce (MYO)Fibroblast Contraction in a New 3D Skin Model to Understand the Inflammation-Fibrosis Axis in Systemic Sclerosis

doi: 10.64898/2026.02.12.705496

Figure Lengend Snippet: ( A ) Sorted CD45-cells (fibroblasts) were analyzed for expression of myofibroblast-related genes after co-culture with monocytes in collagen hydrogels. Values (mean ± SEM) indicate relative gene expression (−ΔCt) measured with qPCR. Reference genes used were GAPDH and RPS27A. Analyses were done by two-way ANOVA with Sidak’s multiple comparisons test (**p<0.01, ***p<0.001, ****p<0.0001). ( B ) Representative immunohistochemistry staining showing robust expression of α-SMA (alpha-smooth muscle actin) (left) and fibroblast activation protein (FAP) (right) in the hydrogels containing only fibroblasts (top) compared to those co-culture with monocytes (bottom). The small blue nuclei in the bottom panels are monocytes. (Myo)fibroblasts are clearly positive for both markers (black arrows), with a typically elongated shape.

Article Snippet: Sections were then incubated with primary rabbit –anti-human alpha-smooth muscle actin (α-SMA) (1:200) (ab5694, Abcam, UK), – fibroblast activation protein (FAP) (1:100) (ab207178, Abcam), – Phospho-SMAD2 (1:100) (#3108L, CellSignaling, USA), – Phospho-Stat3 (1:50) (#9145, CellSignaling), or mouse anti-human – CD68 (#MCA1815, BioRad, USA) (1:200) antibodies, for 1 hour at room temperature.

Techniques: Expressing, Co-Culture Assay, Gene Expression, Immunohistochemistry, Staining, Activation Assay