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mouse gas6 elisa  (R&D Systems)


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    R&D Systems mouse gas6 elisa
    The γ-carboxylated protein <t>GAS6</t> is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM
    Mouse Gas6 Elisa, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+gas6/Mouse+Gas6+DuoSet+ELISA/pmc13121459-207-10-16
    Average 93 stars, based on 15 article reviews
    mouse gas6 elisa - by Bioz Stars, 2026-10
    93/100 stars

    Images

    1) Product Images from "Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice"

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    Journal: Bone Research

    doi: 10.1038/s41413-026-00528-2

    The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM
    Figure Legend Snippet: The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

    Techniques Used: Expressing, Gene Expression, Derivative Assay, Western Blot, Phospho-proteomics

    TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant
    Figure Legend Snippet: TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Techniques Used: Staining, Derivative Assay, Cell Culture, Recombinant

    Gamma-carboxylated GAS6 impacts on osteoclast differentiation and fusion. a–d Gene expression analysis by qPCR of osteoclast differentiation markers Acp5 (TRAP), Clcn7 , Ctsk , and Dcstamp . Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for 2, 4, and 6 days ( n = 4 per condition). e Schematic representation of the assay used to assess the impact of γ-carboxylated GAS6 on pre-osteoclast fusion in culture using a conditionally activated tdTomato (Tom) reporter (created with BioRender). f Representative pictures of live osteoclast cultures at the indicated time and concentration of recombinant γ-carboxylated GAS6. The stars indicate the presence of fusion events (Tom + cells) in presence of GAS6 at Day 4. g Quantification of the number of fusion events per 10 mm 2 at the indicated time of osteoclasts differentiation ( n = 16 fields per condition). Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a – d ) and ( g ). *** P < 0.001, ** P < 0.01
    Figure Legend Snippet: Gamma-carboxylated GAS6 impacts on osteoclast differentiation and fusion. a–d Gene expression analysis by qPCR of osteoclast differentiation markers Acp5 (TRAP), Clcn7 , Ctsk , and Dcstamp . Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for 2, 4, and 6 days ( n = 4 per condition). e Schematic representation of the assay used to assess the impact of γ-carboxylated GAS6 on pre-osteoclast fusion in culture using a conditionally activated tdTomato (Tom) reporter (created with BioRender). f Representative pictures of live osteoclast cultures at the indicated time and concentration of recombinant γ-carboxylated GAS6. The stars indicate the presence of fusion events (Tom + cells) in presence of GAS6 at Day 4. g Quantification of the number of fusion events per 10 mm 2 at the indicated time of osteoclasts differentiation ( n = 16 fields per condition). Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a – d ) and ( g ). *** P < 0.001, ** P < 0.01

    Techniques Used: Gene Expression, Derivative Assay, Cell Culture, Recombinant, Concentration Assay

    GAS6 promotes osteoclast formation and bone resorption in vivo. a–j Six-month-old WT (non-transgenic littermates) and ApoE-Gas6 Tg male mice were analyzed. GAS6 concentration in the serum ( a ) and bone marrow cavity ( b ) ( n = 4). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections ( n = 15–17). e–h μCT analysis of the distal femur trabecular bone ( n = 12). e Representative μCT images. f Quantification of trabecular bone volume (BV/TV). g Quantification of trabecular bone surface density (BS/TV). h Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness, respectively; Conn.Dn., connectivity density. i–j Bone histomorphometry analysis of lumbar vertebrae in six-month-old WT and ApoE-Gas6 Tg male ( n = 19–20). i Representative pictures of TRAP staining. j Number of osteoclasts per bone perimeter (N.Oc/B.Pm) and osteoclast surface over bone surface (Oc.S/BS). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( a , b , d , f – h and j ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant
    Figure Legend Snippet: GAS6 promotes osteoclast formation and bone resorption in vivo. a–j Six-month-old WT (non-transgenic littermates) and ApoE-Gas6 Tg male mice were analyzed. GAS6 concentration in the serum ( a ) and bone marrow cavity ( b ) ( n = 4). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections ( n = 15–17). e–h μCT analysis of the distal femur trabecular bone ( n = 12). e Representative μCT images. f Quantification of trabecular bone volume (BV/TV). g Quantification of trabecular bone surface density (BS/TV). h Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness, respectively; Conn.Dn., connectivity density. i–j Bone histomorphometry analysis of lumbar vertebrae in six-month-old WT and ApoE-Gas6 Tg male ( n = 19–20). i Representative pictures of TRAP staining. j Number of osteoclasts per bone perimeter (N.Oc/B.Pm) and osteoclast surface over bone surface (Oc.S/BS). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( a , b , d , f – h and j ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Techniques Used: In Vivo, Transgenic Assay, Concentration Assay, Staining, Derivative Assay

    Related Articles

    Recombinant:

    Article Title: Cleavage of Mer Tyrosine Kinase (MerTK) from the Cell Surface Contributes to the Regulation of Retinal Phagocytosis
    Article Snippet: .. Recombinant proteins and antibodies against mouse MerTK, mouse MFG-E8, and mouse Gas6 were from R & D Systems, and recombinant protein S was from MP Biomedicals. .. Other antibodies used against various proteins were as follows: PEDF (Abcam); mouse CD51 (αv integrin) and mouse Tyro3 (BD Transduction Laboratories); rabbit β5 integrin H-96, MerTK E-20, and IRBP C-18 (Santa Cruz Biotechnology); rabbit tubulin (Sigma); rabbit ZO-1 (Invitrogen); and rabbit phospho-MerTK (FabGennix Inc. International).

    Article Title: Cleavage of Mer Tyrosine Kinase (MerTK) from the Cell Surface Contributes to the Regulation of Retinal Phagocytosis
    Article Snippet: .. Recombinant proteins and antibodies against mouse MerTK, mouse MFG-E8, and mouse Gas6 were from R & D Systems, and recombinant protein S was from MP Biomedicals. .. Other antibodies used against various proteins were as follows: PEDF (Abcam); mouse CD51 ( v integrin) and mouse Tyro3 (BD Transduction Laboratories); rabbit 5 integrin H-96, MerTK E-20, and IRBP C-18 (Santa Cruz Biotechnology); rabbit tubulin (Sigma); rabbit ZO-1 (Invitrogen); and rabbit phospho-MerTK (FabGennix Inc. International).

    Pore Size:

    Article Title: AXL antibodies
    Article Snippet: .. For this purpose, in the morning of day 1, NIH3T3-AXL cl.7 cells were seeded on 15 cm dished in normal growth medium, which in the evening was replaced by serum-reduced medium (DMEM, 0.5% FCS) in order to starve cells for 36 h. The next day, a FluoroBlock 96 well plate (Becton Dickinson #351164, 8 μm pore size) was coated with 10 μg collagen 1/ml 0.1 M acetic acid over night at 37° C. On day 3, the serum-reduced medium (DMEM, 0.5% FCS) was replaced by serum-free medium (DMEM, 0% FCS, 0.1% BSA) for another 4 h. Cells were harvested with 10 mM EDTA in PBS and pre-incubated with rat anti-AXL antibodies 4A6, 11B7 or 2A1 at a cell density of 4×105 cells/ml and an antibody concentration of 10 μg/ml for 45 min. 50 μl cell suspension (20,000 cells) per well were then placed in the top chamber of the FluoroBlock 96-well plate, 225 μl medium (DMEM, 0% FCS, 0.1% BSA) with or without 400 ng/ml mouse Gas6 (R&D Systems) were used per well in the bottom chamber. .. Cells were left to migrate for 7 h at 37° C. and stained afterwards with 4.2 μM calcein-AM (Molecular Probes #C3099) in PBS, 1 mM CaCl2, 1 mM MgCl2 for 1 h at 37° C. Using a Victor plate reader (Perkin Elmer), the fluorescence of each well was measured at a wavelength of 530 nm.

    Concentration Assay:

    Article Title: AXL antibodies
    Article Snippet: .. For this purpose, in the morning of day 1, NIH3T3-AXL cl.7 cells were seeded on 15 cm dished in normal growth medium, which in the evening was replaced by serum-reduced medium (DMEM, 0.5% FCS) in order to starve cells for 36 h. The next day, a FluoroBlock 96 well plate (Becton Dickinson #351164, 8 μm pore size) was coated with 10 μg collagen 1/ml 0.1 M acetic acid over night at 37° C. On day 3, the serum-reduced medium (DMEM, 0.5% FCS) was replaced by serum-free medium (DMEM, 0% FCS, 0.1% BSA) for another 4 h. Cells were harvested with 10 mM EDTA in PBS and pre-incubated with rat anti-AXL antibodies 4A6, 11B7 or 2A1 at a cell density of 4×105 cells/ml and an antibody concentration of 10 μg/ml for 45 min. 50 μl cell suspension (20,000 cells) per well were then placed in the top chamber of the FluoroBlock 96-well plate, 225 μl medium (DMEM, 0% FCS, 0.1% BSA) with or without 400 ng/ml mouse Gas6 (R&D Systems) were used per well in the bottom chamber. .. Cells were left to migrate for 7 h at 37° C. and stained afterwards with 4.2 μM calcein-AM (Molecular Probes #C3099) in PBS, 1 mM CaCl2, 1 mM MgCl2 for 1 h at 37° C. Using a Victor plate reader (Perkin Elmer), the fluorescence of each well was measured at a wavelength of 530 nm.

    Article Title: MicroRNA-34a dependent regulation of AXL controls the activation of dendritic cells in inflammatory arthritis
    Article Snippet: In the next step, cDNA for AXL, IL-6 and housekeeping 18S were amplified with PreAmp Master kit (#4391128/ThermoFisher Scientific) and cDNA for RNU6, miR-34a were amplified with miScript PreAMP PCR Kit (#331452/Qiagen) followed by standard qPCR as described above. .. The concentration of cytokines and chemokines in mouse and human DC culture supernatants was quantified by immuno-fluorescence assay using a 20-plex cytokine assay (LMC0006; Invitrogen) on a Bio-Plex platform (Bio-Rad) or using paired antibodies ELISAs (mouse and human TNF, #CHC3013 and #CHC1753/Life Technologies; mouse IL-23 (#88723088/ ThermoFisher Scientific) and mouse GAS6 (#DY986/R&D Systems) . .. The data were analysed by Graph Pad Prism version 5.0 and Minitab 17 software.

    Suspension:

    Article Title: AXL antibodies
    Article Snippet: .. For this purpose, in the morning of day 1, NIH3T3-AXL cl.7 cells were seeded on 15 cm dished in normal growth medium, which in the evening was replaced by serum-reduced medium (DMEM, 0.5% FCS) in order to starve cells for 36 h. The next day, a FluoroBlock 96 well plate (Becton Dickinson #351164, 8 μm pore size) was coated with 10 μg collagen 1/ml 0.1 M acetic acid over night at 37° C. On day 3, the serum-reduced medium (DMEM, 0.5% FCS) was replaced by serum-free medium (DMEM, 0% FCS, 0.1% BSA) for another 4 h. Cells were harvested with 10 mM EDTA in PBS and pre-incubated with rat anti-AXL antibodies 4A6, 11B7 or 2A1 at a cell density of 4×105 cells/ml and an antibody concentration of 10 μg/ml for 45 min. 50 μl cell suspension (20,000 cells) per well were then placed in the top chamber of the FluoroBlock 96-well plate, 225 μl medium (DMEM, 0% FCS, 0.1% BSA) with or without 400 ng/ml mouse Gas6 (R&D Systems) were used per well in the bottom chamber. .. Cells were left to migrate for 7 h at 37° C. and stained afterwards with 4.2 μM calcein-AM (Molecular Probes #C3099) in PBS, 1 mM CaCl2, 1 mM MgCl2 for 1 h at 37° C. Using a Victor plate reader (Perkin Elmer), the fluorescence of each well was measured at a wavelength of 530 nm.

    Cytokine Assay:

    Article Title: MicroRNA-34a dependent regulation of AXL controls the activation of dendritic cells in inflammatory arthritis
    Article Snippet: In the next step, cDNA for AXL, IL-6 and housekeeping 18S were amplified with PreAmp Master kit (#4391128/ThermoFisher Scientific) and cDNA for RNU6, miR-34a were amplified with miScript PreAMP PCR Kit (#331452/Qiagen) followed by standard qPCR as described above. .. The concentration of cytokines and chemokines in mouse and human DC culture supernatants was quantified by immuno-fluorescence assay using a 20-plex cytokine assay (LMC0006; Invitrogen) on a Bio-Plex platform (Bio-Rad) or using paired antibodies ELISAs (mouse and human TNF, #CHC3013 and #CHC1753/Life Technologies; mouse IL-23 (#88723088/ ThermoFisher Scientific) and mouse GAS6 (#DY986/R&D Systems) . .. The data were analysed by Graph Pad Prism version 5.0 and Minitab 17 software.

    Control:

    Article Title: AXL antibodies
    Article Snippet: .. The next day, cells were starved in serum-reduced medium (DMEM, 0.5% FCS) for 10 h and subsequently pre-incubated with 20 μg/ml of the mouse control antibody 72A1, the antagonistic rat anti-AXL antibodies 11D5 and 11B7, as well as the agonistic antibody 2A1 in DMEM, 0.5% FCS for 1 h at 37° C. Cells were treated with or without 400 ng/ml mouse Gas6 (R&D Systems) by adding ligand directly to the antibody solution, and were then left to grow for 96 h. AlamarBlueTM (BIOSOURCE #DAL1100) was added and incubated at 37° C. in the dark. ..

    Incubation:

    Article Title: AXL antibodies
    Article Snippet: .. The next day, cells were starved in serum-reduced medium (DMEM, 0.5% FCS) for 10 h and subsequently pre-incubated with 20 μg/ml of the mouse control antibody 72A1, the antagonistic rat anti-AXL antibodies 11D5 and 11B7, as well as the agonistic antibody 2A1 in DMEM, 0.5% FCS for 1 h at 37° C. Cells were treated with or without 400 ng/ml mouse Gas6 (R&D Systems) by adding ligand directly to the antibody solution, and were then left to grow for 96 h. AlamarBlueTM (BIOSOURCE #DAL1100) was added and incubated at 37° C. in the dark. ..



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    R&D Systems recombinant mouse gas6 protein
    Increased bone mass in male mice lacking γ-carboxylation in osteoblasts. a Gene expression analysis by qPCR of Ggcx and Vkorc1 in bone marrow derived monocytes (BMMC), osteoclasts (OCL), proliferating pre-osteoblasts (pre-OB), and mineralized <t>osteoblast</t> (OB) cultures ( n = 3). b Protein expression in liver (Liv) and bone cells by Western blot. GGCX was analyzed on a 7.5% SDS Tris Glycine gel using 20 μg of extracts, while VKORC1 was resolved on a 10% SDS Tris Tricine gel using 10 μg of extracts. c–g Six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice were analyzed ( n = 7–10). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections. e Representative μCT images of the distal femur trabecular bones. f Quantification of trabecular bone volume (BV/TV) and trabecular bone surface density (BS/TV) from the μCT data. g Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness respectively; Conn.Dn., connectivity density. Unpaired, 2-tailed Student’s t test was used in ( d ), ( f ), and ( g ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant
    Recombinant Mouse Gas6 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+gas6/Recombinant+Mouse+Gas6+Protein/pm41885342-31-18-22
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    anti gas6  (R&D Systems)
    94
    R&D Systems anti gas6
    Increased bone mass in male mice lacking γ-carboxylation in osteoblasts. a Gene expression analysis by qPCR of Ggcx and Vkorc1 in bone marrow derived monocytes (BMMC), osteoclasts (OCL), proliferating pre-osteoblasts (pre-OB), and mineralized <t>osteoblast</t> (OB) cultures ( n = 3). b Protein expression in liver (Liv) and bone cells by Western blot. GGCX was analyzed on a 7.5% SDS Tris Glycine gel using 20 μg of extracts, while VKORC1 was resolved on a 10% SDS Tris Tricine gel using 10 μg of extracts. c–g Six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice were analyzed ( n = 7–10). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections. e Representative μCT images of the distal femur trabecular bones. f Quantification of trabecular bone volume (BV/TV) and trabecular bone surface density (BS/TV) from the μCT data. g Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness respectively; Conn.Dn., connectivity density. Unpaired, 2-tailed Student’s t test was used in ( d ), ( f ), and ( g ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant
    Anti Gas6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+gas6/Mouse+Gas6+Antibody/pm41885342-31-9-10
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    anti gas6 - by Bioz Stars, 2026-10
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    mouse gas6  (Elabscience Biotechnology)
    94
    Elabscience Biotechnology mouse gas6
    TAM receptor (TYRO3, AXL, MERTK) and ligand (PROS1, <t>GAS6)</t> 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.
    Mouse Gas6, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+gas6/Mouse+Gas6+(Growth+Arrest+Specific+Protein+6)+ELISA+Kit/pmc13046968-311-0-13
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    p re ss enzyme linked immunosorbent assay elisa mouse gas6  (Elabscience Biotechnology)
    94
    Elabscience Biotechnology p re ss enzyme linked immunosorbent assay elisa mouse gas6
    TAM receptor (TYRO3, AXL, MERTK) and ligand (PROS1, <t>GAS6)</t> 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.
    P Re Ss Enzyme Linked Immunosorbent Assay Elisa Mouse Gas6, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Expressing, Gene Expression, Derivative Assay, Western Blot, Phospho-proteomics

    TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Staining, Derivative Assay, Cell Culture, Recombinant

    Gamma-carboxylated GAS6 impacts on osteoclast differentiation and fusion. a–d Gene expression analysis by qPCR of osteoclast differentiation markers Acp5 (TRAP), Clcn7 , Ctsk , and Dcstamp . Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for 2, 4, and 6 days ( n = 4 per condition). e Schematic representation of the assay used to assess the impact of γ-carboxylated GAS6 on pre-osteoclast fusion in culture using a conditionally activated tdTomato (Tom) reporter (created with BioRender). f Representative pictures of live osteoclast cultures at the indicated time and concentration of recombinant γ-carboxylated GAS6. The stars indicate the presence of fusion events (Tom + cells) in presence of GAS6 at Day 4. g Quantification of the number of fusion events per 10 mm 2 at the indicated time of osteoclasts differentiation ( n = 16 fields per condition). Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a – d ) and ( g ). *** P < 0.001, ** P < 0.01

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: Gamma-carboxylated GAS6 impacts on osteoclast differentiation and fusion. a–d Gene expression analysis by qPCR of osteoclast differentiation markers Acp5 (TRAP), Clcn7 , Ctsk , and Dcstamp . Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for 2, 4, and 6 days ( n = 4 per condition). e Schematic representation of the assay used to assess the impact of γ-carboxylated GAS6 on pre-osteoclast fusion in culture using a conditionally activated tdTomato (Tom) reporter (created with BioRender). f Representative pictures of live osteoclast cultures at the indicated time and concentration of recombinant γ-carboxylated GAS6. The stars indicate the presence of fusion events (Tom + cells) in presence of GAS6 at Day 4. g Quantification of the number of fusion events per 10 mm 2 at the indicated time of osteoclasts differentiation ( n = 16 fields per condition). Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a – d ) and ( g ). *** P < 0.001, ** P < 0.01

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Gene Expression, Derivative Assay, Cell Culture, Recombinant, Concentration Assay

    GAS6 promotes osteoclast formation and bone resorption in vivo. a–j Six-month-old WT (non-transgenic littermates) and ApoE-Gas6 Tg male mice were analyzed. GAS6 concentration in the serum ( a ) and bone marrow cavity ( b ) ( n = 4). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections ( n = 15–17). e–h μCT analysis of the distal femur trabecular bone ( n = 12). e Representative μCT images. f Quantification of trabecular bone volume (BV/TV). g Quantification of trabecular bone surface density (BS/TV). h Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness, respectively; Conn.Dn., connectivity density. i–j Bone histomorphometry analysis of lumbar vertebrae in six-month-old WT and ApoE-Gas6 Tg male ( n = 19–20). i Representative pictures of TRAP staining. j Number of osteoclasts per bone perimeter (N.Oc/B.Pm) and osteoclast surface over bone surface (Oc.S/BS). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( a , b , d , f – h and j ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: GAS6 promotes osteoclast formation and bone resorption in vivo. a–j Six-month-old WT (non-transgenic littermates) and ApoE-Gas6 Tg male mice were analyzed. GAS6 concentration in the serum ( a ) and bone marrow cavity ( b ) ( n = 4). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections ( n = 15–17). e–h μCT analysis of the distal femur trabecular bone ( n = 12). e Representative μCT images. f Quantification of trabecular bone volume (BV/TV). g Quantification of trabecular bone surface density (BS/TV). h Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness, respectively; Conn.Dn., connectivity density. i–j Bone histomorphometry analysis of lumbar vertebrae in six-month-old WT and ApoE-Gas6 Tg male ( n = 19–20). i Representative pictures of TRAP staining. j Number of osteoclasts per bone perimeter (N.Oc/B.Pm) and osteoclast surface over bone surface (Oc.S/BS). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( a , b , d , f – h and j ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: In Vivo, Transgenic Assay, Concentration Assay, Staining, Derivative Assay

    IL4@CHHSSSARC-EV promoted microglial phagocytosis via GAS6-MERTK pathway. A Representative immunofluorescence staining images of IBA1 and C-C3 on retinal flat mounts from OIR mice treated with PBS or IL4@CHHSSSARC-EV. Scale bar: 100 μm. Enlarged: 50 μm. B qPCR analysis of GAS6 and TAM systems (Axl, MERTK, Tyro3) mRNA levels in M1 microglia exposed to PBS or IL4@CHHSSSARC-EV ( n = 3). C Concentrations of GAS6 in the supernatant of M1 microglia after exposed to PBS or IL4@CHHSSSARC-EV by ELISA ( n = 3). D Representative western blot images and quantitative analysis of protein expression of p-MERTK and MERTK after M1 microglia were exposed to PBS or IL4@CHHSSSARC-EV ( n = 3). E Representative immunofluorescence staining images of IBA1 and C-C3 on retinal flat mounts from OIR mice treated with IL4@CHHSSSARC-EV or IL4@CHHSSSARC-EV plus UNC2025. Scale bar: 100 μm. Enlarged: 50 μm. F Representative western blot images and quantitative analysis of protein expression of p-MERTK and MERTK after M1 microglia were exposed to IL4@CHHSSSARC-EV or IL4@CHHSSSARC-EV plus UNC2025 ( n = 3). G Representative immunofluorescence staining images of IB4 in retinas from OIR mice treated with IL4@CHHSSSARC-EV or IL4@CHHSSSARC-EV plus UNC2025 ( n = 5). Scale bar: 500 μm. White region indicated neovascular area and yellow-circulated region marked avascular area. H Quantitative analysis of neovascular area and avascular area of G . * P < 0.1, ** P < 0.01, *** P < 0.001

    Journal: Journal of Nanobiotechnology

    Article Title: Microglia-specific interleukin-4 delivery by engineered extracellular vesicles restores inner blood-retinal barrier in diabetic retinopathy via GAS6-MERTK pathway

    doi: 10.1186/s12951-025-03976-w

    Figure Lengend Snippet: IL4@CHHSSSARC-EV promoted microglial phagocytosis via GAS6-MERTK pathway. A Representative immunofluorescence staining images of IBA1 and C-C3 on retinal flat mounts from OIR mice treated with PBS or IL4@CHHSSSARC-EV. Scale bar: 100 μm. Enlarged: 50 μm. B qPCR analysis of GAS6 and TAM systems (Axl, MERTK, Tyro3) mRNA levels in M1 microglia exposed to PBS or IL4@CHHSSSARC-EV ( n = 3). C Concentrations of GAS6 in the supernatant of M1 microglia after exposed to PBS or IL4@CHHSSSARC-EV by ELISA ( n = 3). D Representative western blot images and quantitative analysis of protein expression of p-MERTK and MERTK after M1 microglia were exposed to PBS or IL4@CHHSSSARC-EV ( n = 3). E Representative immunofluorescence staining images of IBA1 and C-C3 on retinal flat mounts from OIR mice treated with IL4@CHHSSSARC-EV or IL4@CHHSSSARC-EV plus UNC2025. Scale bar: 100 μm. Enlarged: 50 μm. F Representative western blot images and quantitative analysis of protein expression of p-MERTK and MERTK after M1 microglia were exposed to IL4@CHHSSSARC-EV or IL4@CHHSSSARC-EV plus UNC2025 ( n = 3). G Representative immunofluorescence staining images of IB4 in retinas from OIR mice treated with IL4@CHHSSSARC-EV or IL4@CHHSSSARC-EV plus UNC2025 ( n = 5). Scale bar: 500 μm. White region indicated neovascular area and yellow-circulated region marked avascular area. H Quantitative analysis of neovascular area and avascular area of G . * P < 0.1, ** P < 0.01, *** P < 0.001

    Article Snippet: The concentration of GAS6 was determined by the mouse GAS6 ELISA kit (EK2384, Multi Sciences).

    Techniques: Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

    Increased bone mass in male mice lacking γ-carboxylation in osteoblasts. a Gene expression analysis by qPCR of Ggcx and Vkorc1 in bone marrow derived monocytes (BMMC), osteoclasts (OCL), proliferating pre-osteoblasts (pre-OB), and mineralized osteoblast (OB) cultures ( n = 3). b Protein expression in liver (Liv) and bone cells by Western blot. GGCX was analyzed on a 7.5% SDS Tris Glycine gel using 20 μg of extracts, while VKORC1 was resolved on a 10% SDS Tris Tricine gel using 10 μg of extracts. c–g Six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice were analyzed ( n = 7–10). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections. e Representative μCT images of the distal femur trabecular bones. f Quantification of trabecular bone volume (BV/TV) and trabecular bone surface density (BS/TV) from the μCT data. g Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness respectively; Conn.Dn., connectivity density. Unpaired, 2-tailed Student’s t test was used in ( d ), ( f ), and ( g ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: Increased bone mass in male mice lacking γ-carboxylation in osteoblasts. a Gene expression analysis by qPCR of Ggcx and Vkorc1 in bone marrow derived monocytes (BMMC), osteoclasts (OCL), proliferating pre-osteoblasts (pre-OB), and mineralized osteoblast (OB) cultures ( n = 3). b Protein expression in liver (Liv) and bone cells by Western blot. GGCX was analyzed on a 7.5% SDS Tris Glycine gel using 20 μg of extracts, while VKORC1 was resolved on a 10% SDS Tris Tricine gel using 10 μg of extracts. c–g Six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice were analyzed ( n = 7–10). c Representative pictures of sections from lumbar vertebrae stained with von Kossa and van Gieson. d Quantification of trabecular bone volume over tissue volume (BV/TV) from the L4 and L5 lumbar vertebrae sections. e Representative μCT images of the distal femur trabecular bones. f Quantification of trabecular bone volume (BV/TV) and trabecular bone surface density (BS/TV) from the μCT data. g Trabecular bone µCT derived data. Tb.Sp., Tb.N., and Tb.Th., trabecular spacing, number, and thickness respectively; Conn.Dn., connectivity density. Unpaired, 2-tailed Student’s t test was used in ( d ), ( f ), and ( g ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Gene Expression, Derivative Assay, Expressing, Western Blot, Staining

    Reduced osteoclast number and surface in Ggcx ff ;OCN-Cre male mice. a–h Bone histomorphometry analysis of lumbar vertebrae in six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice ( n = 6–10). a Representative pictures of calcein double labeling and toluidine blue staining. b Mineral apposition rate (MAR). c Bone formation rate over bone surface (BFR/BS). d Number of osteoblasts per bone perimeter (N.Ob/B.Pm). e Osteoblast surface over bone surface (Ob.S/BS). f Representative pictures of TRAP staining. g Number of osteoclasts per bone perimeter (N.Oc/B.Pm). h Osteoclast surface over bone surface (Oc.S/BS). i Fasting serum CTx levels ( n = 12–17). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( b – e ) and ( g – i ). ** P < 0.01, * P < 0.05, ns: non-significant

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: Reduced osteoclast number and surface in Ggcx ff ;OCN-Cre male mice. a–h Bone histomorphometry analysis of lumbar vertebrae in six-month-old Ggcx ff and Ggcx ff ;OCN-Cre male mice ( n = 6–10). a Representative pictures of calcein double labeling and toluidine blue staining. b Mineral apposition rate (MAR). c Bone formation rate over bone surface (BFR/BS). d Number of osteoblasts per bone perimeter (N.Ob/B.Pm). e Osteoblast surface over bone surface (Ob.S/BS). f Representative pictures of TRAP staining. g Number of osteoclasts per bone perimeter (N.Oc/B.Pm). h Osteoclast surface over bone surface (Oc.S/BS). i Fasting serum CTx levels ( n = 12–17). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( b – e ) and ( g – i ). ** P < 0.01, * P < 0.05, ns: non-significant

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Labeling, Staining

    Ggcx inactivation impairs the ability of osteoblasts to support osteoclastogenesis ex vivo. a Representative TRAP staining of osteoblasts (OB) and bone marrow cells (BM) co-cultures at day 8 in the presence of prostaglandin E 2 (PGE 2 ; 10 –6 mol/L) and 1,25 vitamin D 3 (VitD 3 ; 10 –8 mol/L). Ggcx ff osteoblasts were transduced with either Ad-GFP (control) or Ad-Cre (knockout) before the addition of the WT bone marrow cells. b Quantification of the number of TRAP + osteoclasts per well (Nb of OCL/well) ( n = 3). c Representative TRAP staining of osteoblasts and bone marrow cells co-cultures at day 8. Control ( Ocn + / + ) or osteocalcin-deficient ( Ocn -/- ) osteoblasts were cultured with WT bone marrow cells. d Quantification of the number of TRAP + osteoclasts per well (Nb of OCL/well) ( n = 3). e Gene expression analysis by qPCR in Ggcx ff + Ad-GFP and Ggcx ff + Ad-Cre osteoblasts cultured in presence (+) or absence (–) of PGE 2 and VitD 3 for 6 days. Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( b ) and ( d ). Two-way ANOVA with Bonferroni’s posttests was used in ( e ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: Ggcx inactivation impairs the ability of osteoblasts to support osteoclastogenesis ex vivo. a Representative TRAP staining of osteoblasts (OB) and bone marrow cells (BM) co-cultures at day 8 in the presence of prostaglandin E 2 (PGE 2 ; 10 –6 mol/L) and 1,25 vitamin D 3 (VitD 3 ; 10 –8 mol/L). Ggcx ff osteoblasts were transduced with either Ad-GFP (control) or Ad-Cre (knockout) before the addition of the WT bone marrow cells. b Quantification of the number of TRAP + osteoclasts per well (Nb of OCL/well) ( n = 3). c Representative TRAP staining of osteoblasts and bone marrow cells co-cultures at day 8. Control ( Ocn + / + ) or osteocalcin-deficient ( Ocn -/- ) osteoblasts were cultured with WT bone marrow cells. d Quantification of the number of TRAP + osteoclasts per well (Nb of OCL/well) ( n = 3). e Gene expression analysis by qPCR in Ggcx ff + Ad-GFP and Ggcx ff + Ad-Cre osteoblasts cultured in presence (+) or absence (–) of PGE 2 and VitD 3 for 6 days. Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in ( b ) and ( d ). Two-way ANOVA with Bonferroni’s posttests was used in ( e ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Ex Vivo, Staining, Transduction, Control, Knock-Out, Cell Culture, Gene Expression

    The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: The γ-carboxylated protein GAS6 is expressed by osteoblasts and activates its receptors on pre-osteoclasts. a Expression analysis by qPCR of genes encoding the known γ-carboxylated proteins. Gas6 is highlighted in red. b Gene expression analysis by qPCR of the TAM receptors Axl , Mertk and Tyro3 in bone marrow derived monocytes (d0: day 0) and in differentiating osteoclast cultures in the presence of RANKL and M-CSF (d2-d6: day 2 to 6; n = 4). c Western blot analysis of the phosphorylation (P) of AKT (S473) and TAM (Y702 in AXL and Y753 in MerTK) in bone marrow derived monocytes (BMMC) serum starved for 3 h and treated with GAS6 (200 ng/mL) for the indicated times. Total AKT, MerTK, and AXL were used as loading controls. d Quantification of P-AKT (S473), P-MerTK (Y753), and P-AXL (Y702) normalized to the amount of total protein ( n = 3). Results represent the mean ± SEM

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Expressing, Gene Expression, Derivative Assay, Western Blot, Phospho-proteomics

    TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Journal: Bone Research

    Article Title: Vitamin K-dependent carboxylation in osteoblasts regulates bone resorption through GAS6 in male mice

    doi: 10.1038/s41413-026-00528-2

    Figure Lengend Snippet: TAM signaling and γ-carboxylated GAS6 signaling promotes osteoclast formation in culture. Representative TRAP staining (left) and quantification of the TRAP + osteoclast area (right) in WT osteoblast (OB) and bone marrow cell (BM) co-cultures at day 8 in the presence of PGE 2 and VitD 3 , with or without the TAM inhibitors LDC1267 ( a ) or R428 ( b ), at the indicated concentrations. c–f Bone marrow derived monocytes (BMMC) were cultured in the presence of RANKL (20 ng/mL) and M-CSF (10 ng/mL) with or without recombinant γ-carboxylated GAS6 at the indicated concentrations for up to 6 days. c Representative TRAP staining at day 5 and 6 of differentiation. d Quantification of the TRAP + osteoclast area at day 4, 5, and 6 of differentiation. e Quantification of the number of TRAP + multinucleated osteoclasts at day 4, 5, and 6 of differentiation. f Quantification of the number of nuclei per osteoclast at day 5 of differentiation. Results represent the mean ± SEM. One-way ANOVA with Bonferroni’s posttests was used in ( a , b ) and ( d – f ). *** P < 0.001, ** P < 0.01, * P < 0.05, ns: non-significant

    Article Snippet: GAS6 from serum and osteoblast supernatant were quantified using a mouse GAS6 ELISA (DuoSet ELISA DY986, R&D Systems, Minneapolis, MN) as we previously described.

    Techniques: Staining, Derivative Assay, Cell Culture, Recombinant

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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