Journal: Biomolecules

Article Title: Glycogen Hydrogel Loaded with Schistosoma japonicas Peptide SJMHE1 Improves Skin Wound Healing

doi: 10.3390/biom16030392

Figure Lengend Snippet: Macrophages treated with SJMHE1 promote fibroblast and HUVEC migration through TGF-β1/Smad3 pathway and VEGFA expression. ( A ) TGF-β1 protein expression in RAW264.7 cells assessed by Western blotting. ( B ) p-Smad3 protein expression in L929cells assessed by Western blotting. ( C ) Relative mRNA expression of COL1a1 in L929 cells. ( D ) Cell scratch assay and number of migrating L929 cells (scale bar = 200 μm). ( E ) p-Smad3 protein expression in HUVECs assessed by Western blotting. ( F ) Cell scratch assay and migration of HUVECs (scale bar = 200 μm). ( G ) VEGFA expression in THP-1-induced macrophages. ( H ) Representative images of vascular network formation and tube length quantification in HUVECs in vitro (scale bar = 200 μm). Data are presented as mean ± SD ( n = 3). ns = no significance, * p < 0.05, ** p < 0.01, and *** p < 0.001. Original Western blot images are provided in the .

Article Snippet: After washing with PBS, L929 cells and HUVECs were treated with the following low-serum (2% FBS) conditioned media: CM-Control, CM-SJMHE1, CM-SJMHE1 supplemented with 5 μg/mL TGF-β1 neutralising antibody (BE0057, Bio X Cell, Lebanon, PA, USA), or normal medium supplemented with 10 ng/mL TGF-β1 recombinant protein (HY- P70648 , MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Migration, Expressing, Western Blot, Wound Healing Assay, In Vitro

Journal: Biomolecules

Article Title: Glycogen Hydrogel Loaded with Schistosoma japonicas Peptide SJMHE1 Improves Skin Wound Healing

doi: 10.3390/biom16030392

Figure Lengend Snippet: SJMHE1-gel treatment increases TGF-β1, p-Smad3, and VEGFA expression in mice. ( A ) Representative immunohistochemical staining images of TGF-β1, p-Smad3, and VEGFA on day 7 (scale bar = 50 μm). ( B ) Quantification of TGF-β1, p-Smad3 ( C ) and VEGFA ( D ) expression. Data are presented as mean ± SD ( n = 6). ns = no significance, * p < 0.05, ** p < 0.01.

Article Snippet: After washing with PBS, L929 cells and HUVECs were treated with the following low-serum (2% FBS) conditioned media: CM-Control, CM-SJMHE1, CM-SJMHE1 supplemented with 5 μg/mL TGF-β1 neutralising antibody (BE0057, Bio X Cell, Lebanon, PA, USA), or normal medium supplemented with 10 ng/mL TGF-β1 recombinant protein (HY- P70648 , MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Wound Repair and Regeneration

Article Title: Dendritic Cell–Associated MARCKSL1 Regulates Fibroblast Differentiation During Wound Healing

doi: 10.1111/wrr.70143

Figure Lengend Snippet: Induction of MARCKSL1 expression in mouse DCs by wounding stimulation. (A) Immunofluorescence staining of DCs following TGF‐β1 stimulation. Red: MARCKSL1; Green: CD11c; Blue: DAPI. Scale bar = 10 μm. (B) Western blot analysis of MARCKSL1 and p‐MARCKS expression in TGF‐β1‐treated DCs. * p < 0.05. (C) RT‐qPCR analysis of MARCKSL1 mRNA expression in TGF‐β1‐stimulated DCs. * p < 0.05. Each experiment was independently repeated three times (biological replicates). Within each experiment, three technical replicates were performed per condition and averaged prior to statistical analysis. Each data point represents one independent biological replicate. Data are shown as mean ± SEM. DAPI, 4′,6‐diamidino‐2‐phenylindole; DCs, dendritic cells; MARCKSL1, myristoylated alanine‐rich C‐kinase substrate‐like 1; TGF‐β1, transforming growth factor beta 1; CD11c, cluster of differentiation 11c.

Article Snippet: Cells were subsequently stimulated with 5 ng/mL TGF‐β1 (7666‐MB‐005, R&D Systems, Minneapolis, MN, USA) for 24 h. Following stimulation, DCs were washed thrice with phosphate‐buffered saline (PBS) to remove residual cytokines and resuspended in fresh medium.

Techniques: Expressing, Immunofluorescence, Staining, Western Blot, Quantitative RT-PCR

Journal: Wound Repair and Regeneration

Article Title: Dendritic Cell–Associated MARCKSL1 Regulates Fibroblast Differentiation During Wound Healing

doi: 10.1111/wrr.70143

Figure Lengend Snippet: MARCKSL1‐positive DCs promote fibroblast activation. (A) Immunofluorescence staining of co‐cultures of TGF‐β1‐stimulated DCs and fibroblasts. Green: Phalloidin; Red: CD11c; Blue: α‐SMA; White: MARCKSL1; Cyan: DAPI. Scale bar = 10 μm. (B) Western blot analysis of α‐SMA expression in fibroblasts after co‐culture with TGF‐β1‐stimulated DCs. * p < 0.05. (C) RT‐qPCR analysis of Acta2 (α‐SMA) expression in fibroblasts after co‐culture with TGF‐β1‐stimulated DCs. * p < 0.05. (D) Validation of MARCKSL1 knockdown in DCs transfected with pLKO.1‐shMARCKSL1 under TGF‐β1 stimulation. * p < 0.05. (E) Immunofluorescence staining of fibroblast co‐cultures with MARCKSL1‐shRNA‐treated DCs. Green: Phalloidin; Red: CD11c; Blue: α‐SMA; White: MARCKSL1; Cyan: DAPI. Scale bar = 10 μm. (F) Western blot analysis of α‐SMA expression in fibroblasts after co‐culture with MARCKSL1‐shRNA‐treated DCs. * p < 0.05. (G) RT‐qPCR analysis of Acta2 expression in fibroblasts co‐cultured with MARCKSL1‐shRNA‐treated DCs. * p < 0.05. Each experiment was independently repeated three times (biological replicates). Within each experiment, three technical replicates were performed per condition and averaged prior to statistical analysis. Each data point represents one independent biological replicate. Data are shown as mean ± SEM. CD11c, cluster of differentiation 11c; DAPI, 4′,6‐diamidino‐2‐phenylindole; DCs, dendritic cells; MARCKSL1, myristoylated alanine‐rich C‐kinase substrate‐like 1; RT‐qPCR, real‐time quantitative polymerase chain reaction; TGF‐β1, transforming growth factor beta 1; α‐SMA, α‐smooth muscle actin.

Article Snippet: Cells were subsequently stimulated with 5 ng/mL TGF‐β1 (7666‐MB‐005, R&D Systems, Minneapolis, MN, USA) for 24 h. Following stimulation, DCs were washed thrice with phosphate‐buffered saline (PBS) to remove residual cytokines and resuspended in fresh medium.

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Expressing, Co-Culture Assay, Quantitative RT-PCR, Biomarker Discovery, Knockdown, Transfection, shRNA, Cell Culture, Real-time Polymerase Chain Reaction

Journal: Journal of Biomedical Science

Article Title: Intestinal epithelial Smad7 drives purine metabolic dysregulation and ileal inflammation

doi: 10.1186/s12929-026-01224-3

Figure Lengend Snippet: TGF-β1 enhances CD73 expression in ileal epithelial cells. a . Representative Western blots of phosphorylated Smad3 (p-Smad3) and β-actin in IECs isolated from the ileum of Smad7Tg mice (n = 1) and stimulated or not with recombinant murine TGF-β1 (20 ng/ml) for 15 and 30 min. b. Expression of Nt5e RNA transcripts in intestinal epithelial cells isolated from the ileum of Smad7Tg mice (n = 5) and stimulated or not with recombinant murine TGF-β1 (20 ng/ml) for 1.5 and 3 h. Data are presented as mean ± SEM, with each point representing the expression of Nt5e in a single sample. Differences among groups were compared using one-way analysis of variance (ANOVA) followed by the Tukey test (** P ≤ .01, *** P ≤ .001). Ut: untreated

Article Snippet: IECs isolated from control mice were stimulated with recombinant murine TGF-β1 (20 ng/ml, #7666, R&D Systems, Minneapolis, Minnesota, USA) or adenosine (1 mM, #A4036, Sigma) for 1.5 or 3 h and then collected and resuspended in lysis buffer supplemented with 1% β-mercaptoethanol and stored at -80 °C until RNA and protein extraction.

Techniques: Expressing, Western Blot, Isolation, Recombinant