Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: TAM receptor (TYRO3, AXL, MERTK) and ligand (PROS1, GAS6) expression in: a keloid-prone ( n = 8, K) versus healthy ( n = 6, N) individual wounds ( GSE113619 ; 1st and 2nd: initial and 42-day biopsies); b normal human skin ( n = 24, NS), early (≤7 days, n = 22, EW), late (8 days–1 month, n = 29, LW), and chronic (>1 month, n = 3, CW) wounds ( GSE178411 ); c normal ( n = 5, NS), acute ( n = 5, AW), type I ( n = 5, DW Ⅰ), and type II diabetic ( n = 3, DW Ⅱ) mouse wounds ( GSE283228 ); d IFN-γ/LPS-stimulated macrophages ( n = 4; GSE53986 ). Conserved GAS6 downregulation in datasets e GSE113619 , f GSE178411 , g GSE283228 , and h GSE53986 (mean ± SD; n values as in a – d ). i – l Single-cell RNA-seq analysis of human wounds ( GSE241132 ; n = 3). i UMAP plot colored by cell type. j GAS6 expression over time. k FeaturePlot visualization of GAS6 expression at representative times. l GAS6 expression across cell types. m – o In vivo analysis of RU-301-treated mice. m Representative wound images (POD0, 3, 7, 10, 14) and H&E-stained scar (POD30); scale bars, 1 mm, 500 µm. n Relative wound area over time (Ctrl, n = 6; RU-301, n = 7). o Scar length on POD30 (Ctrl, n = 3; RU-301, n = 3). p , r Apoptosis in POD3 wounds. p Quantification of Cleaved caspase 3 MFI (Control, n = 6; RU-301, n = 5). r Representative immunofluorescence images; scale bars, 500 µm, 100 µm. q , s Macrophage migration towards POD3 dermal samples. q Quantification of migrated macrophages ( n = 3). s Representative Transwell images; scale bars, 100 µm. All quantitative data are presented as mean ± SD. Statistical significance was determined using two-tailed unpaired t-tests for comparisons between two groups or one-way ANOVA with Tukey’s post-hoc test for multi-group comparisons. n represents independent biological experiments. Ctrl control group, POD postoperative day, MFI mean fluorescence intensity. Control groups received vehicle. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Expressing, Single Cell, RNA Sequencing, In Vivo, Staining, Control, Immunofluorescence, Migration, Two Tailed Test, Fluorescence

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a Schematic of LNP-GAS6 mRNA design for targeted TAM receptor agonism. b – g Physicochemical characterization of LNPs and LNP-GAS6 mRNA. b – d Diameter, polydispersity index (PDI), and zeta potential measured by dynamic light scattering (DLS) ( n = 5). Encapsulation efficiency ( e ) and encapsulated mRNA concentration ( f ) quantified using the Quant-iT RiboGreen RNA Assay Kit ( n = 5). g Representative transmission electron microscopy (TEM) image; scale bars, 200 nm, 50 nm. h GAS6 protein secretion quantified by ELISA in fibroblasts and macrophages at 24, 48, and 72 h post-treatment with LNP-GAS6 mRNA ( n = 3). i – k Intracellular GAS6 protein expression assessed by immunofluorescence. i Representative images of fibroblasts and macrophages; Blue, DAPI (nuclei); green, GAS6; scale bars, 100 µm. Quantification of GAS6 MFI in fibroblasts ( j ) and macrophages ( k ) ( n = 3). All quantitative data are presented as mean ± SD. Statistical significance was determined using two-tailed unpaired t-tests for comparisons between two groups or one-way ANOVA with Tukey’s post-hoc test for multi-group comparisons. n represents independent biological experiments. MFI mean fluorescence intensity; I, LNP; II, LNP-GAS6 mRNA. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Zeta Potential Analyzer, Encapsulation, Concentration Assay, Transmission Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Two Tailed Test, Fluorescence

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a Il10 expression in macrophages after GAS6 siRNA treatment, assessed by RT‑qPCR ( n = 3). Expression of Csf1 ( b ) and Cxcl12 ( c ) in fibroblasts treated with conditioned medium (CM) from the indicated macrophage groups ( n = 3). d Percentage of macrophages that engulfed apoptotic cells (CFDA‑SE⁺), assessed by flow cytometry ( n = 3). Expression of Nos2 ( e ) and Arg1 ( f ) in macrophages ( n = 3). g – i Efferocytosis analysis in macrophages. g Percentage of CFDA‑SE⁺ macrophages. h MFI of CFDA‑SE within phagocytes, reflecting the number of apoptotic cells ingested per macrophage ( n = 4). i Representative flow cytometry plots. j Schematic of the macrophage‑fibroblast paracrine signaling model. Created in BioRender. Hanxiao, S. (2026) https://BioRender.com/snsu6jk . Expression of Tnf ( k ), Il1b ( l ), and Il10 ( m ) in macrophages ( n = 3). Expression of Csf1 ( n ) and Cxcl12 ( o ) in fibroblasts ( n = 3). LPS CM: conditioned medium from LPS‑treated macrophages; LNP‑GAS6 mRNA CM: conditioned medium from LNP‑GAS6 mRNA‑treated macrophages. Expression of Csf1 ( p ) and Cxcl12 ( q ) in fibroblasts under the indicated treatments ( n = 3). All quantitative data are presented as mean ± SD. Statistical significance was determined using two‑tailed unpaired t‑tests for comparisons between two groups or one‑way ANOVA with Tukey’s post‑hoc test for multi‑group comparisons. CM conditioned medium, MFI mean fluorescence intensity. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a , b Thermosensitive properties of LNP-GAS6 mRNA@Gel. a In vitro sol‑gel transition in vitro at 4 °C (liquid) and 37 °C (gel) (representative images, n = 3). b In vivo gelation upon application to acute and diabetic murine wounds (representative images, n = 3). c Distribution of PKH26‑labeled LNPs within the hydrogel matrix, detected by confocal microscopy (representative image, n = 3). d – f Bioluminescence imaging of luciferase expression. d Representative images of wounds after treatment with LNP‑Fluc mRNA (5 μg/wound) by injection or hydrogel. e Longitudinal quantification of bioluminescence flux in individual mice over time ( n = 4). f Comparative analysis of total signal intensity between LNP‑Fluc mRNA and LNP‑Fluc mRNA@Gel groups ( n = 4). g – k In vivo experiment of LNP-mRNA@Hydrogel concentration screening. g Representative immunofluorescence images for GAS6 (green) and DAPI (blue) in POD3 acute wounds; scale bars, 1 mm. Quantification of GAS6 MFI in deep and superficial skin layers at the wound center ( h ) and edge ( i ) ( n = 4). Wound width ( j ) and granulation tissue thickness ( k ) after treatment ( n = 4). All quantitative data are presented as mean ± SD. Statistical significance was determined using two‑tailed unpaired t‑tests for comparisons between two groups or one‑way ANOVA with Tukey’s post‑hoc test for multi‑group comparisons. n represents independent biological experiments. MFI mean fluorescence intensity, POD postoperative day. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: In Vitro, In Vivo, Confocal Microscopy, Imaging, Luciferase, Expressing, Injection, Concentration Assay, Immunofluorescence, Fluorescence

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a Experimental design: full-thickness wounds in mice treated with PBS, Gel, LNP@Gel, LNP-GAS6 mRNA@Gel, or GAS6 recombinant protein. Created in BioRender. Hanxiao, S. (2026) https://BioRender.com/78012r9 . b – d Wound closure analysis. b Representative wound images on POD0, 3, 7, 10, 14, and 30. c Quantification of relative wound area over time ( n = 8). d Heatmap of wound closure dynamics ( n = 8). e – g Scar analysis on POD30. e Representative H&E (scale bars, 1 mm, 200 µm) and Masson’s trichrome (scale bars, 1 mm, 250 µm, 50 µm) staining. f Scar length ( n = 6). g CVF ratio (scar area CVF/normal skin CVF) ( n = 6). h , i Apoptosis in POD3 wounds. ( h ) Representative images of immunofluorescence for Cleaved caspase 3; scale bars, 200 µm, 100 µm. i Quantification of Cleaved caspase 3 MFI ( n = 3). Blue, DAPI; Green, F4/80; Red, Cleaved Caspase 3. j , k Macrophage migration toward POD3 dermal samples ( n = 3). j Representative images of Transwell assay; scale bars, 100 µm. k Quantification of migrated macrophages ( n = 3). All quantitative data are presented as mean ± SD. Statistical significance was determined using two‑tailed unpaired t‑tests for comparisons between two groups or one‑way ANOVA with Tukey’s post‑hoc test for multi‑group comparisons. n represents independent biological experiments. CVF collagen volume fraction, MFI mean fluorescence intensity, POD postoperative day. Ⅰ, Control; Ⅱ, Gel; Ⅲ, LNP@Gel, Ⅳ, LNP-GAS6 mRNA@Gel; Ⅴ, GAS6 Recombinant Protein. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Recombinant, Staining, Immunofluorescence, Migration, Transwell Assay, Fluorescence, Control

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a Administration protocol for LNP-GAS6 mRNA@Gel with or without AXL or MERTK receptor inhibitors. b , c Wound closure analysis. b Quantification of relative wound area over time ( n = 4). c Representative wound images on POD0, 3, 7, 10, and 14. d – h Transcriptomic analysis of treatment effects ( n = 3). d KEGG and e GSEA enrichment plots highlighting OXPHOS as a top-ranked pathway. Venn diagram ( f ), heatmap ( g ), and volcano plot ( h ) showing expression changes of OXPHOS-related genes (45 upregulated, 3 downregulated). i – l OXPHOS functional validation in macrophages ( n = 3). Expression of Ndufa3 ( i ), Uqcrb ( j ), and Cox5a ( k ) by RT‑qPCR following GAS6 siRNA treatment ( n = 3). l Relative ATP levels in macrophages after treatment with GAS6 siRNA or LNP‑GAS6 mRNA. All quantitative data are presented as mean ± SD. Statistical significance was determined using two‑tailed unpaired t‑tests for comparisons between two groups or one‑way ANOVA with Tukey’s post‑hoc test for multi‑group comparisons. n represents independent biological experiments. Ctrl control, AXL i AXL inhibitor, MERTK i MERTK inhibitor, OXPHOS oxidative phosphorylation, POD postoperative day. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Expressing, Functional Assay, Biomarker Discovery, Control, Phospho-proteomics

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a Experimental design: full-thickness wounds on rabbit ears treated with PBS, Gel, LNP@Gel, or LNP-GAS6 mRNA@Gel. Created in BioRender. Hanxiao, S. (2026) https://BioRender.com/eh3yqdy . b Representative wound images on POD0, 3, 7, 14, 21, 28, and 60. c , e Hypertrophic scar assessment on POD60 by Masson’s trichrome staining. c Representative images, each containing scar and adjacent normal skin ( n = 6); scale bars, 1 mm, 500 µm. e Quantification of the SEI, calculated as scar height/normal skin height ( n = 6). d , f Collagen composition analysis on POD60 by Sirius red staining under polarized light. d Representative images (scar and normal skin) ( n = 3); scale bars, 200 µm, 20 µm. f Quantification of the collagen I/III ratio ( n = 3). Under polarized light, thick type I collagen fibers appear red/orange-red; thin type III collagen fibers appear green. All quantitative data are presented as mean ± SD. Statistical significance was determined using two‑tailed unpaired t‑tests for comparisons between two groups or one‑way ANOVA with Tukey’s post‑hoc test for multi‑group comparisons. n represents independent biological experiments. SEI scar elevation index, POD postoperative day. Ⅰ, Control; Ⅱ, Gel; Ⅲ, LNP@Gel, Ⅳ, LNP-GAS6 mRNA@Gel; Ⅴ, Normal Skin. Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Staining, Control

Journal: Nature Communications

Article Title: Spatiotemporally controlled restoration of GAS6 signaling via mRNA therapy promotes scarless healing in preclinical models

doi: 10.1038/s41467-026-69540-8

Figure Lengend Snippet: a Experimental design: full-thickness dorsal wounds treated with Gel, LNP@Gel, LNP-GAS6 mRNA@Gel, or human EGF gel. Created in BioRender. Hanxiao, S. (2026) https://BioRender.com/a0axfcg . b – e Wound closure analysis. b Representative wound images on POD0, 7, 14, 21, 28, and 35. c Quantification of relative wound area over time ( n = 6). d Heatmap of closure dynamics ( n = 6). e Schematic simulation of wound area changes during healing. f – h Scar analysis on POD14 and 35. f Representative Masson’s trichrome staining; scale bars, 1 mm, 200 µm. g Scar length quantification ( n = 6). h CVF ratio (scar area CVF/normal skin CVF) ( n = 6). All quantitative data are presented as mean ± SD. Statistical significance was determined using two‑tailed unpaired t‑tests for comparisons between two groups or one‑way ANOVA with Tukey’s post‑hoc test for multi‑group comparisons. n represents independent biological experiments. CVF collagen volume fraction, POD postoperative day. Ⅰ, Gel; Ⅱ, LNP@Gel; Ⅲ, LNP-GAS6 mRNA@Gel, Ⅳ, h-EGF Gel (Human-Epidermal Growth Factor Gel). Source data are provided as a file.

Article Snippet: Mouse GAS6 (#E-EL-M0282), IL-6 (#E-EL-M0044), and TNF-α (#E-EL-M3063) ELISA Kit was purchased from Elabscience.

Techniques: Staining