Journal: bioRxiv

Article Title: Senescence-induced reparative fibroblasts enable scarless wound healing in aged murine skin

doi: 10.1101/2025.04.17.648896

Figure Lengend Snippet: (A) Schematic diagram of study design, multi-omics analysis, and validation cohort. (B) Hematoxylin-eosin (H&E) and Masson staining for scar tissues, displaying morphological differences between the young and aged groups. Scale bar, 200 um. (C) Immunofluorescence staining for Sox9 (marker of the bulge) and Ki67 (marker of de novo HF) in the scar tissues of the aged group. Scale bar, 50um. (D) Quantification analysis for numbers of de novo HFs (n=5 per group). Significances were analyzed using bidirectional t-tests. (E-F) Diagram of ECM ultrastructure analysis(E). TSNE plots of unsupervised cluster analysis of ECM characteristics in each group with representative Sirius red staining images. Scale bar, 50um. (F-H) Bulk RNA sequencing for scar tissues at 28 dpw(n=3 per group). Gene ontology enrichment analysis showing the top biological process terms based on differentially expressed genes (p<0.05, |log2FC|>1)(G). Gene set enrichment analysis (GSEA) showing regeneration-associated terms (left) and heatmap of corresponding genes (right)(H). Abbreviation: dpw, day post wound. HF, hair follicle. Epi, epidermis. ECM, extracellular matrix. TSNE, T-Distributed Stochastic Neighbor Embedding. EGFR, epidermal growth factor receptor. Statistical thresholds followed p = 0.05 convention (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001), with nonsignificant (ns) determinations requiring p > 0.05.

Article Snippet: Ki-67( ) and SOX9( ) were utilized for observation of hair follicle, while CRABP1(12588-1-AP, proteintech), a-SMA(67735-1-Ig, proteintech), PRSS35(GTX123037, GeneTex), PTN(HA722055, HUABIO), and EREG(ER65835, HUABIO) were employed to assess the fibroblast subcluster.

Techniques: Biomarker Discovery, Staining, Immunofluorescence, Marker, RNA Sequencing

Journal: bioRxiv

Article Title: Senescence-induced reparative fibroblasts enable scarless wound healing in aged murine skin

doi: 10.1101/2025.04.17.648896

Figure Lengend Snippet: (A) Schematic diagram showing treatment design in vivo and vitro assays. (B)Hematoxylin-eosin (H&E) and Masson staining for scar tissues of control, PTN, and EREG treatment samples. Scale bar, 200 um. (C-D) Immunofluorescence staining for Sox9 (yellow) and Ki67 (purple) in the scar tissues of the EREG treatment group. Dotted lines signed morphology of de novo HF. Scale bar, 50um (C). Quantification analysis for numbers of de novo HFs (n=5 per group). Significances were utilized using bidirectional t-tests (D). (E) TSNE plots of unsupervised cluster analysis of ECM characteristics in each group with representative Sirius red staining images. Scale bar, 100um. (F) Immunofluorescence staining for CRABP1 (green, marker of papilla fibroblast), and PRSS35 (red, marker of reparative fibroblast) in the scar tissues. Scale bar, 50um(left). Quantification analysis for the ratio of marker protein area/DAPI area (n=5 per group). Significances were utilized using bidirectional t-tests(middle). Colocation plot showing colocalization of CRABP1/PRSS35 assessed by Pearson correlation coefficient(right). (G) Immunofluorescence staining for F-actin (red, cytoskeleton) and a-SMA (green, marker of fibroblast activation) in murine fibroblast lineage (L929 cells). Scale bar as shown in images. The difference in mean fluorescence intensity was utilized by bidirectional t-tests (n=5). (H-I) TSNE plots of unsupervised cluster analysis of ECM characteristics in the EREG group with representative Sirius red staining images. Scale bar, 100um(H). Quantification analysis for the ratio of collagen deposition area in scar tissues assessed by bidirectional t-tests (n=5). Abbreviation: HF, hair follicle. Epi, epidermis. EREG_R, EREG_regeneration area. EREG_S, EREG_scar area.a-SMA, a-smooth muscle actin. P value (*p < 0.05; **p < 0.01; ***p < 0.001), with nonsignificant (ns) determinations requiring p > 0.05.

Article Snippet: Ki-67( ) and SOX9( ) were utilized for observation of hair follicle, while CRABP1(12588-1-AP, proteintech), a-SMA(67735-1-Ig, proteintech), PRSS35(GTX123037, GeneTex), PTN(HA722055, HUABIO), and EREG(ER65835, HUABIO) were employed to assess the fibroblast subcluster.

Techniques: In Vivo, Staining, Control, Immunofluorescence, Marker, Activation Assay, Fluorescence